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TW202444421A - Immunogenic compositions comprising conjugated capsular saccharide antigens and uses thereof - Google Patents

Immunogenic compositions comprising conjugated capsular saccharide antigens and uses thereof Download PDF

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TW202444421A
TW202444421A TW113111933A TW113111933A TW202444421A TW 202444421 A TW202444421 A TW 202444421A TW 113111933 A TW113111933 A TW 113111933A TW 113111933 A TW113111933 A TW 113111933A TW 202444421 A TW202444421 A TW 202444421A
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考希克 杜塔
凱特林 高拉格
伊西斯 坎恩夫斯基
進煥 金
賈斯汀 凱斯 摩蘭
蘇德漢 辛格
艾柏席賽克 拉文卓 法塔克
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美商輝瑞大藥廠
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Abstract

The present invention relates to new immunogenic compositions comprising conjugated Streptococcus pneumoniae capsular saccharide antigens (glycoconjugates), kits comprising said immunogenic compositions and uses thereof. Immunogenic compositions of the present invention will typically comprise at least one glycoconjugate from a S. pneumoniae serotype not found in PREVNAR<SP>®</SP>, SYNFLORIX<SP>®</SP> and/or PREVNAR 13<SP>®</SP>. The invention also relates to vaccination of human subjects, in particular infants and elderly, against pneumococcal infections using said novel immunogenic compositions.

Description

包含經結合之莢膜醣抗原的免疫原組合物及其用途Immunogenic compositions comprising bound capsular glycogen antigens and uses thereof

本發明係關於免疫原組合物及疫苗之領域、其製造及此類組合物在醫學中之用途。The present invention relates to the field of immunogenic compositions and vaccines, their manufacture and the use of such compositions in medicine.

更特定言之，其係關於經分離之肺炎鏈球菌( Streptococcus pneumoniae)血清型38醣、其醣結合物、用於製造肺炎鏈球菌血清型38醣結合物之方法及包含肺炎鏈球菌血清型38醣結合物之免疫原組合物。 More specifically, it relates to isolated Streptococcus pneumoniae serotype 38 saccharides, saccharide conjugates thereof, methods for producing Streptococcus pneumoniae serotype 38 saccharide conjugates, and immunogenic compositions comprising Streptococcus pneumoniae serotype 38 saccharide conjugates.

本發明亦關於分析經分離之肺炎鏈球菌血清型38多醣、經還原之血清型38多醣或肺炎鏈球菌血清型38醣結合物之分析方法。The present invention also relates to a method for analyzing isolated S. pneumoniae serotype 38 polysaccharide, reduced S. pneumoniae serotype 38 polysaccharide or S. pneumoniae serotype 38 saccharide conjugate.

本發明之肺炎鏈球菌血清型38醣及醣結合物可用作疫苗。The Streptococcus pneumoniae serotype 38 saccharide and saccharide conjugate of the present invention can be used as a vaccine.

數十年來，已成功地利用藉由將此等分子結合至「載體」分子來提高弱免疫原分子之免疫原性的方法(參見例如Goebel等人(1939) J. Exp. Med. 69: 53)。舉例而言，已描述了許多免疫原組合物，其中經純化之莢膜聚合物已與載體蛋白結合，以藉由採用此「載體效應」來產生更高效的免疫原組合物。(Schneerson等人(1984) Infect. Immun. 45: 582-591)。亦已證明，結合可避免在用游離多醣進行免疫接種時通常在嬰兒中觀測到的不良抗體反應(Anderson等人(1985) J. Pediatr. 107: 346；Insel等人(1986) J. Exp. Med. 158: 294)。For decades, methods have been successfully used to increase the immunogenicity of weak immunogenic molecules by conjugating such molecules to "carrier" molecules (see, e.g., Goebel et al. (1939) J. Exp. Med. 69: 53). For example, a number of immunogenic compositions have been described in which purified capsular polymers have been conjugated to carrier proteins to produce more potent immunogenic compositions by exploiting this "carrier effect." (Schneerson et al. (1984) Infect. Immun. 45: 582-591). Conjugation has also been shown to avoid the adverse antibody reactions commonly observed in infants when immunized with free polysaccharides (Anderson et al. (1985) J. Pediatr. 107: 346; Insel et al. (1986) J. Exp. Med. 158: 294).

已成功使用各種交聯劑或偶合劑，諸如同型雙官能、異型雙官能或零長度交聯劑來產生結合物。目前許多方法可供用於將免疫原分子(諸如醣、蛋白質及肽)與肽或蛋白質載體偶合。大部分方法產生胺、醯胺、胺基甲酸酯、異硫脲或二硫鍵，或在一些情況下產生硫醚。使用將反應性位點引入載體及/或免疫原分子上之反應性胺基酸分子之側鏈中的交聯劑或偶合劑的不足之處在於，反應性位點在未中和的情況下會在活體外與任何非所需分子自由反應(因此可能不利地影響結合物之功能性或穩定性)，或在活體內與任何非所需分子自由反應(因此在用製劑進行免疫接種之個人或動物中引起不良事件的可能風險)。可利用各種已知的化學反應來使此類過量的反應性位點發生反應或對其進行「封端」，以便使此等位點不活化，但此等反應可能在其他方面破壞結合物之功能性。A variety of crosslinkers or coupling agents, such as isobifunctional, heterobifunctional, or zero-length crosslinkers, have been used successfully to produce conjugates. Many methods are currently available for coupling immunogenic molecules (such as carbohydrates, proteins, and peptides) to peptide or protein carriers. Most methods produce amine, amide, carbamate, isothiourea, or disulfide bonds, or in some cases thioethers. A disadvantage of using cross-linkers or coupling agents that introduce reactive sites into the side chains of reactive amino acid molecules on the carrier and/or immunogenic molecules is that the reactive sites, if unneutralized, are free to react with any undesired molecules in vitro (thus potentially adversely affecting the functionality or stability of the conjugate), or in vivo (thus presenting a possible risk of causing adverse events in individuals or animals immunized with the formulation). Various known chemical reactions can be used to react or "block" such excess reactive sites in order to render them inactive, but such reactions may otherwise disrupt the functionality of the conjugate.

因此，仍需要新的經適當封端之醣結合物及製備該等結合物之方法，使得功能性得以保留，且結合物仍能夠誘發所需免疫反應。Therefore, there remains a need for new appropriately capped carbohydrate conjugates and methods of preparing such conjugates such that functionality is retained and the conjugates are still able to induce a desired immune response.

肺炎鏈球菌多醣，特別係莢膜多醣，係在細菌表面上發現的重要免疫原。此已導致其成為肺炎鏈球菌疫苗設計中之重要組分。已證實莢膜多醣尤其在連接至載體蛋白時可用於誘發免疫反應。Pneumococcal polysaccharides, particularly capsular polysaccharides, are important immunogens found on the surface of the bacteria. This has led to them becoming important components in the design of pneumococcal vaccines. Capsular polysaccharides have been shown to be useful in eliciting an immune response, particularly when linked to a carrier protein.

因此，需要能夠對肺炎鏈球菌血清型38產生穩健免疫反應之抗原。Therefore, there is a need for antigens that can generate a robust immune response to S. pneumoniae serotype 38.

本發明尤其提供肺炎鏈球菌血清型38醣結合物。The present invention particularly provides Streptococcus pneumoniae serotype 38 saccharide conjugates.

為了滿足此等及其他需求，本發明係關於一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目，其中Y表示絲胺酸或甘胺酸殘基且其中所有該等重複單元包含相同的Y殘基。 To meet these and other needs, the present invention relates to an isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: wherein n represents the number of repeating units, wherein Y represents a serine or glycine residue and wherein all of the repeating units contain the same Y residue.

在一態樣中，本發明係關於一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目，其中Y表示絲胺酸或甘胺酸殘基，其中所有該等重複單元包含相同的Y殘基，且其中在β-D-Gal p4OAc,6(Y)之位置4處的O-乙醯基存在於約0%至約100%的該等重複單元中。在一實施例中，該O-乙醯基存在於約50%至約100%的該等重複單元中。較佳地，該O-乙醯基存在於約80%至約100%的該等重複單元中。甚至更佳地，該O-乙醯基存在於約90%至約100%的該等重複單元中。 In one aspect, the present invention relates to an isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: wherein n represents the number of repeating units, wherein Y represents a serine or glycine residue, wherein all of the repeating units comprise the same Y residue, and wherein the O-acetyl group at position 4 of β-D-Gal p 4OAc,6(Y) is present in about 0% to about 100% of the repeating units. In one embodiment, the O-acetyl group is present in about 50% to about 100% of the repeating units. Preferably, the O-acetyl group is present in about 80% to about 100% of the repeating units. Even more preferably, the O-acetyl group is present in about 90% to about 100% of the repeating units.

在某些實施例中，本發明之經分離之肺炎鏈球菌血清型38醣具有10至5,000個重複單元。In certain embodiments, the isolated S. pneumoniae serotype 38 saccharide of the present invention has 10 to 5,000 repeat units.

在一態樣中，本發明提供一種醣結合物，其由與載體蛋白結合的具有以上所揭露之重複單元的醣組成。In one aspect, the present invention provides a carbohydrate conjugate, which is composed of a carbohydrate having the above-disclosed repeating units conjugated to a carrier protein.

本發明進一步提供帶有還原之α-D-2-乙醯胺基-2,6-二去氧基-木糖-己-4-酮糖(α-D-Sug)殘基之肺炎鏈球菌血清型38醣結合物。The present invention further provides a Streptococcus pneumoniae serotype 38 saccharide conjugate having a reduced α-D-2-acetamido-2,6-dideoxy-xylose-hex-4-ketose (α-D-Sug) residue.

在一態樣中，本發明係關於此類結合物之製備模式。In one aspect, the present invention relates to a method for preparing such a conjugate.

在一實施例中，本發明係關於一種包含本發明之肺炎鏈球菌血清型38醣及/或本發明之肺炎鏈球菌血清型38醣結合物之免疫原組合物。In one embodiment, the present invention relates to an immunogenic composition comprising the Streptococcus pneumoniae serotype 38 saccharide of the present invention and/or the Streptococcus pneumoniae serotype 38 saccharide conjugate of the present invention.

本發明進一步關於肺炎鏈球菌血清型38醣及結合物之分析方法。The present invention further relates to a method for analyzing sugars and conjugates of Streptococcus pneumoniae serotype 38.

本發明部分地基於藉由使用NMR光譜法鑑別新穎的肺炎鏈球菌多醣結構。咸信本文中所提供之結構為肺炎鏈球菌血清型38的第一次鑑別或第一次正確鑑別。The present invention is based in part on the identification of novel pneumococcal polysaccharide structures by using NMR spectroscopy. It is believed that the structure provided herein is the first identification or the first correct identification of pneumococcal serotype 38.

所產生之(及純化之)多醣用於產生多醣-蛋白結合物(醣結合物)。肺炎鏈球菌血清型38具有獨特多醣結構，其導致在設計結合物生產過程時的獨特考慮。本發明之肺炎鏈球菌血清型38醣結合物亦具有獨特結構及設計。The produced (and purified) polysaccharides are used to produce polysaccharide-protein conjugates (glycoconjugates). S. pneumoniae serotype 38 has a unique polysaccharide structure, which leads to unique considerations in designing the conjugate production process. The S. pneumoniae serotype 38 glycoconjugates of the present invention also have a unique structure and design.

1. 本發明之經分離之肺炎鏈球菌血清型 38 醣如本文所用，與關於醣之術語「分離」係指使用此項技術中已知之純化技術自經純化之多醣分離肺炎鏈球菌血清型特異性莢膜多醣，該等技術包括使用離心、深層過濾、沉澱、超濾、用活性碳處理、透濾及/或管柱層析。一般而言，經分離之多醣係指蛋白質、核酸及非特異性內源性多醣(C-多醣)的部分移除。經分離之多醣含有少於10%、8%、6%、4%或2%蛋白質雜質及/或核酸。經分離之多醣相對於類型特異性多醣含有少於20%之C-多醣。 1. Separated Streptococcus pneumoniae serotype 38 saccharides of the present invention As used herein, the term "separation" with respect to saccharides refers to the separation of Streptococcus pneumoniae serotype-specific capsule polysaccharides from purified polysaccharides using purification techniques known in the art, including the use of centrifugation, deep filtration, precipitation, ultrafiltration, treatment with activated carbon, filtration and/or column chromatography. Generally speaking, separated polysaccharides refer to partial removal of proteins, nucleic acids and non-specific endogenous polysaccharides (C-polysaccharides). Separated polysaccharides contain less than 10%, 8%, 6%, 4% or 2% protein impurities and/or nucleic acids. The isolated polysaccharides contained less than 20% C-polysaccharides relative to the type-specific polysaccharides.

在整個本說明書中，術語「醣」可指示多醣或寡醣且包括兩者。在較佳實施例中，醣為多醣，尤其肺炎鏈球菌血清型38莢膜多醣。Throughout this specification, the term "sugar" may refer to polysaccharides or oligosaccharides and includes both. In a preferred embodiment, the sugar is a polysaccharide, in particular Streptococcus pneumoniae serotype 38 capsular polysaccharide.

在本說明書通篇，術語「血清型38醣」「血清型38莢膜醣」、「肺炎鏈球菌血清型38醣」係指肺炎鏈球菌血清型38莢膜醣且可在本文中互換使用。Throughout this specification, the terms "serotype 38 saccharide", "serotype 38 capsular saccharide", and "Streptococcus pneumoniae serotype 38 capsular saccharide" refer to Streptococcus pneumoniae serotype 38 capsular saccharide and can be used interchangeably herein.

肺炎鏈球菌血清型38莢膜多醣之結構首次揭露且展示於圖1A及圖1B中。本發明人發現血清型38多醣為五醣：α-D-葡糖胺(A)、α-D-2-乙醯胺基-2,6-二去氧基-木糖-己-4-酮糖(α-D-Sug) (B)、O-乙醯化β-D-半乳糖(C)、α-D-半乳糖(D)及β-D-呋喃半乳糖(E)。殘基C (O-乙醯化β-D-半乳糖)在位置6處進一步與絲胺酸或甘胺酸胺基酸連接，由此表示多醣之血清型38絲胺酸及甘胺酸形式。The structure of the pneumococcal serotype 38 capsular polysaccharide was first disclosed and shown in Figures 1A and 1B. The inventors found that the serotype 38 polysaccharide is a pentasaccharide: α-D-glucosamine (A), α-D-2-acetamido-2,6-dideoxy-xylose-hex-4-ketose (α-D-Sug) (B), O-acetylated β-D-galactose (C), α-D-galactose (D) and β-D-furanose (E). The residual C (O-acetylated β-D-galactose) is further linked to a serine or glycine amino acid at position 6, thereby representing the serotype 38 serine and glycine forms of the polysaccharide.

本發明人已發現，肺炎鏈球菌血清型38之某些菌株產生絲胺酸胺基酸連接於Gal p4OAc糖的莢膜多醣(圖1A)，而其他菌株形成甘胺酸附接於Galp4OAc糖的多醣(圖1B)。在表徵血清型38之若干菌株之後，已顯示絲胺酸形式為較豐富的形式。 The inventors have discovered that certain strains of S. pneumoniae serotype 38 produce capsular polysaccharides with serine amino acids linked to Gal p 4OAc sugars ( FIG. 1A ), while other strains form polysaccharides with glycine attached to Gal p 4OAc sugars ( FIG. 1B ). Upon characterization of several strains of serotype 38, the serine form has been shown to be the more abundant form.

因此，在一個實施例中，本發明提供一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目，其中Y表示絲胺酸或甘胺酸殘基且其中所有該等重複單元包含相同的Y殘基。 Therefore, in one embodiment, the present invention provides an isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: wherein n represents the number of repeating units, wherein Y represents a serine or glycine residue and wherein all of the repeating units contain the same Y residue.

因此，在一個實施例中，本發明提供一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目。 Therefore, in one embodiment, the present invention provides an isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: Where n represents the number of repeating units.

在另一實施例中，本發明提供一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目。 In another embodiment, the present invention provides an isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: Where n represents the number of repeating units.

如圖1A及圖1B所示，血清型38多醣之β-D-Gal p4OAc,6Serr(或Gly) (殘基C)在碳位置4經O-乙醯化。O-乙醯化水平可因菌株而異。此外，天然肺炎鏈球菌血清型38醣可例如藉由用鹼(鹼性pH)處理而去乙醯化。如實例部分中所示，天然血清型38多醣可在室溫下用0.25 N NH 4OH培育24小時之後完全去乙醯化。 As shown in Figures 1A and 1B, β-D-Gal p 4OAc,6Serr (or Gly) (residue C) of serotype 38 polysaccharide is O-acetylated at carbon position 4. The level of O-acetylation may vary from strain to strain. In addition, native S. pneumoniae serotype 38 saccharide can be deacetylated, for example, by treatment with a base (alkaline pH). As shown in the Examples section, native serotype 38 polysaccharide can be completely deacetylated after incubation with 0.25 N NH 4 OH at room temperature for 24 hours.

因此，在一個實施例中，本發明提供一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目，其中Y表示絲胺酸或甘胺酸殘基，其中所有該等重複單元包含相同的Y殘基，且其中在β-D-Gal p4OAc,6(Y)之位置4處的O-乙醯基存在於約0%至約100%的該等重複單元中。在一實施例中，該O-乙醯基存在於約50%至約100%的該等重複單元中。較佳地，該O-乙醯基存在於約80%至約100%的該等重複單元中。甚至更佳地，該O-乙醯基存在於約90%至約100%的該等重複單元中。 Therefore, in one embodiment, the present invention provides an isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: wherein n represents the number of repeating units, wherein Y represents a serine or glycine residue, wherein all of the repeating units comprise the same Y residue, and wherein the O-acetyl group at position 4 of β-D-Gal p 4OAc,6(Y) is present in about 0% to about 100% of the repeating units. In one embodiment, the O-acetyl group is present in about 50% to about 100% of the repeating units. Preferably, the O-acetyl group is present in about 80% to about 100% of the repeating units. Even more preferably, the O-acetyl group is present in about 90% to about 100% of the repeating units.

在一個實施例中，本發明提供一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目，其中Y表示絲胺酸或甘胺酸殘基，其中所有該等重複單元包含相同的Y殘基，且其中在β-D-Gal p4OAc,6(Y)之位置4處的O-乙醯基存在於約100%的該等重複單元中。 In one embodiment, the present invention provides an isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: wherein n represents the number of repeating units, wherein Y represents a serine or glycine residue, wherein all of the repeating units comprise the same Y residue, and wherein the O-acetyl group at position 4 of β-D-Gal p 4OAc,6(Y) is present in about 100% of the repeating units.

在一個實施例中，本發明提供一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目，其中Y表示絲胺酸或甘胺酸殘基，其中所有該等重複單元包含相同的Y殘基，且其中在β-D-Gal p4OAc,6(Y)之位置4處的O-乙醯基存在於約95%的該等重複單元中。 In one embodiment, the present invention provides an isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: wherein n represents the number of repeating units, wherein Y represents a serine or glycine residue, wherein all of the repeating units comprise the same Y residue, and wherein the O-acetyl group at position 4 of β-D-Gal p 4OAc,6(Y) is present in about 95% of the repeating units.

因此，在一個實施例中，本發明提供一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目且其中在β-D-Gal p4OAc,6Ser之位置4處的O-乙醯基存在於約0%至約100%的該等重複單元中。在一實施例中，該O-乙醯基存在於約50%至約100%的該等重複單元中。較佳地，該O-乙醯基存在於約80%至約100%的該等重複單元中。甚至更佳地，該O-乙醯基存在於約90%至約100%的該等重複單元中。 Therefore, in one embodiment, the present invention provides an isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: wherein n represents the number of repeating units and wherein the O-acetyl group at position 4 of β-D-Gal p 4OAc,6Ser is present in about 0% to about 100% of the repeating units. In one embodiment, the O-acetyl group is present in about 50% to about 100% of the repeating units. Preferably, the O-acetyl group is present in about 80% to about 100% of the repeating units. Even more preferably, the O-acetyl group is present in about 90% to about 100% of the repeating units.

在一個實施例中，本發明提供一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目且其中在β-D-Gal p4OAc,6Ser之位置4處的O-乙醯基存在於約100%的該等重複單元中。 In one embodiment, the present invention provides an isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: wherein n represents the number of repeating units and wherein the O-acetyl group at position 4 of β-D-Gal p 4OAc,6Ser is present in approximately 100% of the repeating units.

在一個特定實施例中，本發明提供一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目且其中在β-D-Gal p4OAc,6Ser之位置4處的O-乙醯基存在於約95%的該等重複單元中。 In a specific embodiment, the present invention provides an isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: wherein n represents the number of repeating units and wherein the O-acetyl group at position 4 of β-D-Gal p 4OAc,6Ser is present in approximately 95% of the repeating units.

在另一實施例中，本發明提供一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目且其中在β-D-Gal p4OAc,6Gly之位置4處的O-乙醯基存在於約0%至約100%的該等重複單元中。在一實施例中，該O-乙醯基存在於約50%至約100%的該等重複單元中。較佳地，該O-乙醯基存在於約80%至約100%的該等重複單元中。甚至更佳地，該O-乙醯基存在於約90%至約100%的該等重複單元中。 In another embodiment, the present invention provides an isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: wherein n represents the number of repeating units and wherein the O-acetyl group at position 4 of β-D-Gal p 4OAc,6Gly is present in about 0% to about 100% of the repeating units. In one embodiment, the O-acetyl group is present in about 50% to about 100% of the repeating units. Preferably, the O-acetyl group is present in about 80% to about 100% of the repeating units. Even more preferably, the O-acetyl group is present in about 90% to about 100% of the repeating units.

在另一實施例中，本發明提供一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目且其中在β-D-Gal p4OAc,6Gly之位置4處的O-乙醯基存在於約100%的該等重複單元中。 In another embodiment, the present invention provides an isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: wherein n represents the number of repeating units and wherein the O-acetyl group at position 4 of β-D-Gal p 4OAc,6Gly is present in approximately 100% of the repeating units.

在一特定實施例中，本發明提供一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目且其中在β-D-Gal p4OAc,6Gly之位置4處的O-乙醯基存在於95%的該等重複單元中。 In a specific embodiment, the present invention provides an isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: wherein n represents the number of repeating units and wherein the O-acetyl group at position 4 of β-D-Gal p 4OAc,6Gly is present in 95% of the repeating units.

在一實施例中，本發明之肺炎鏈球菌血清型38醣在β-D-Gal p4OAc,6Ser(或Gly)殘基之碳位置4處不帶有O-乙醯基。因此，在一個實施例中，本發明提供一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目，其中Y表示絲胺酸或甘胺酸殘基且其中所有該等重複單元包含相同的Y殘基。 In one embodiment, the pneumococcal serotype 38 saccharide of the present invention does not have an O-acetyl group at carbon position 4 of the β-D-Gal p 4OAc, 6Ser (or Gly) residue. Therefore, in one embodiment, the present invention provides an isolated pneumococcal serotype 38 saccharide having the following repeating units: wherein n represents the number of repeating units, wherein Y represents a serine or glycine residue and wherein all of the repeating units contain the same Y residue.

在一個實施例中，本發明提供一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目。 In one embodiment, the present invention provides an isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: Where n represents the number of repeating units.

在另一實施例中，本發明提供一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目。 In another embodiment, the present invention provides an isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: Where n represents the number of repeating units.

如上文所提及，血清型38醣之β-D-Gal p4OAc,6Ser(或Gly)殘基(殘基C)可在碳位置4處經O-乙醯化。醣中之O-乙醯化水平可不為100%且當醣之結構在此位置處具有由O-乙醯基表示之重複單元時，不應理解為此類醣始終在醣之β-D-Gal p4OAc,6Ser(或Gly)殘基之每一位置4處帶有O-乙醯基。實情為，此表明此等殘基中的大部分經O-乙醯化，較佳地，該O-乙醯基存在於約80%至約100%的該等重複單元中。甚至更佳地，該O-乙醯基存在於約90%至約100%的該等重複單元中。 As mentioned above, the β-D-Gal p 4OAc, 6Ser (or Gly) residue (residue C) of serotype 38 saccharides may be O-acetylated at carbon position 4. The level of O-acetylation in a saccharide may not be 100% and when the structure of a saccharide has repeating units represented by an O-acetyl group at this position, it should not be understood that such saccharides always carry an O-acetyl group at every position 4 of the β-D-Gal p 4OAc, 6Ser (or Gly) residue of the saccharide. Rather, this indicates that the majority of these residues are O-acetylated, preferably, the O-acetyl group is present in about 80% to about 100% of these repeating units. Even more preferably, the O-acetyl group is present in about 90% to about 100% of the repeating units.

在某些實施例中，本發明之經分離之肺炎鏈球菌血清型38醣具有10至5,000個重複單元。在某些態樣中，經分離之醣具有50至4,500個重複單元。在某些態樣中，經分離之醣具有100至4,500個重複單元。在某些態樣中，經分離之醣具有150至2,000個重複單元。In certain embodiments, the isolated S. pneumoniae serotype 38 saccharides of the present invention have 10 to 5,000 repeat units. In certain aspects, the isolated saccharides have 50 to 4,500 repeat units. In certain aspects, the isolated saccharides have 100 to 4,500 repeat units. In certain aspects, the isolated saccharides have 150 to 2,000 repeat units.

來自肺炎鏈球菌血清型38之經分離的莢膜醣可藉由一般熟習此項技術者已知之標準技術來製備。通常，藉由使肺炎鏈球菌血清型38菌株在培養基中(例如在基於大豆之培養基中)生長來產生莢膜多醣，隨後由細菌培養物製備多醣。血清型38肺炎鏈球菌菌株可自已建立的菌種保存中心(例如，疾病控制與預防中心之鏈球菌參考實驗室(Atlanta, GA))或臨床樣本獲得。Isolated capsular polysaccharides from S. pneumoniae serotype 38 can be prepared by standard techniques known to those of ordinary skill in the art. Typically, capsular polysaccharides are produced by growing S. pneumoniae serotype 38 strains in a culture medium, such as a soy-based medium, followed by preparation of the polysaccharide from the bacterial culture. Serotype 38 S. pneumoniae strains can be obtained from established culture collections, such as the Streptococcus Reference Laboratory of the Centers for Disease Control and Prevention (Atlanta, GA), or from clinical specimens.

生物體群體(肺炎鏈球菌血清型38)通常自接種小瓶按比例擴大至接種瓶，且通過一或多個增加體積之接種醱酵器繼代，直至達到生產規模醱酵體積。在生長週期結束時，細胞經溶解且隨後收穫溶解物培養液以供下游(純化)處理(參見例如，WO 2006/110381及WO 2008/118752，美國專利申請公開案第2006/0228380、2006/0228381、2008/0102498及US2008/0286838號)。多醣通常透過離心、沉澱、超濾及/或管柱層析純化(參見例如，WO 2006/110352、WO 2008/118752及WO2020/170190)。The population of organisms (S. pneumoniae serotype 38) is typically scaled up from inoculation vial to inoculation vial and passed through one or more inoculation fermenters of increasing size until a production scale fermentation volume is reached. At the end of the growth cycle, the cells are lysed and the lysate culture is then harvested for downstream (purification) processing (see, e.g., WO 2006/110381 and WO 2008/118752, U.S. Patent Application Publication Nos. 2006/0228380, 2006/0228381, 2008/0102498, and US2008/0286838). Polysaccharides are typically purified by centrifugation, precipitation, ultrafiltration and/or column chromatography (see, e.g., WO 2006/110352, WO 2008/118752 and WO 2020/170190).

經分離之多醣可由不同參數表徵，包括例如重量平均分子量(Mw)。多醣之分子量可藉由尺寸排阻層析(SEC)與多角度雷射光散射偵測器(MALLS)之組合來量測。The isolated polysaccharides can be characterized by various parameters including, for example, weight average molecular weight (Mw). The molecular weight of polysaccharides can be measured by a combination of size exclusion chromatography (SEC) and multi-angle laser light scattering detector (MALLS).

在一實施例中，本發明之經分離之肺炎鏈球菌血清型38醣之重量平均分子量在5 kDa與5000 kDa之間。在一實施例中，經分離之肺炎鏈球菌血清型38醣之重量平均分子量在5 kDa與2000 kDa之間。In one embodiment, the weight average molecular weight of the isolated pneumococcal serotype 38 saccharide of the present invention is between 5 kDa and 5000 kDa. In one embodiment, the weight average molecular weight of the isolated pneumococcal serotype 38 saccharide is between 5 kDa and 2000 kDa.

在一實施例中，經分離之肺炎鏈球菌血清型38多醣之重量平均分子量在50 kDa與5000 kDa之間。在一實施例中，經分離之肺炎鏈球菌血清型38多醣之重量平均分子量在50 kDa與2000 kDa之間。在一實施例中，經分離之肺炎鏈球菌血清型38多醣之重量平均分子量在50 kDa與1000 kDa之間。In one embodiment, the weight average molecular weight of the isolated pneumococcal serotype 38 polysaccharide is between 50 kDa and 5000 kDa. In one embodiment, the weight average molecular weight of the isolated pneumococcal serotype 38 polysaccharide is between 50 kDa and 2000 kDa. In one embodiment, the weight average molecular weight of the isolated pneumococcal serotype 38 polysaccharide is between 50 kDa and 1000 kDa.

在一實施例中，經分離之肺炎鏈球菌血清型38多醣之重量平均分子量在100 kDa與5000 kDa之間。在一實施例中，經分離之肺炎鏈球菌血清型38多醣之重量平均分子量在100 kDa與2000 kDa之間。在一實施例中，經分離之肺炎鏈球菌血清型38多醣之重量平均分子量在100 kDa與1000 kDa之間。在一實施例中，經分離之肺炎鏈球菌血清型38多醣之重量平均分子量在100 kDa與500 kDa之間。In one embodiment, the weight average molecular weight of the isolated pneumococcal serotype 38 polysaccharide is between 100 kDa and 5000 kDa. In one embodiment, the weight average molecular weight of the isolated pneumococcal serotype 38 polysaccharide is between 100 kDa and 2000 kDa. In one embodiment, the weight average molecular weight of the isolated pneumococcal serotype 38 polysaccharide is between 100 kDa and 1000 kDa. In one embodiment, the weight average molecular weight of the isolated pneumococcal serotype 38 polysaccharide is between 100 kDa and 500 kDa.

在一實施例中，經分離之肺炎鏈球菌血清型38多醣之重量平均分子量在300 kDa與5000 kDa之間。在一實施例中，經分離之肺炎鏈球菌血清型38多醣之重量平均分子量在300 kDa與2000 kDa之間。在一實施例中，經分離之肺炎鏈球菌血清型38多醣之重量平均分子量在300 kDa與1000 kDa之間。In one embodiment, the weight average molecular weight of the isolated pneumococcal serotype 38 polysaccharide is between 300 kDa and 5000 kDa. In one embodiment, the weight average molecular weight of the isolated pneumococcal serotype 38 polysaccharide is between 300 kDa and 2000 kDa. In one embodiment, the weight average molecular weight of the isolated pneumococcal serotype 38 polysaccharide is between 300 kDa and 1000 kDa.

在一實施例中，經分離之肺炎鏈球菌血清型38多醣之重量平均分子量在500 kDa與3000 kDa之間。在一實施例中，經分離之多醣之重量平均分子量在500 kDa與2000 kDa之間。在一實施例中，經分離之多醣之重量平均分子量在500 kDa與1000 kDa之間。In one embodiment, the weight average molecular weight of the isolated S. pneumoniae serotype 38 polysaccharide is between 500 kDa and 3000 kDa. In one embodiment, the weight average molecular weight of the isolated polysaccharide is between 500 kDa and 2000 kDa. In one embodiment, the weight average molecular weight of the isolated polysaccharide is between 500 kDa and 1000 kDa.

較佳地，為了產生具有有利可過濾性特性、免疫原性及/或產率之醣結合物，在與載體蛋白結合之前將醣之尺寸設定至目標分子量範圍。Preferably, in order to produce glycoconjugates with favorable filterability properties, immunogenicity and/or yield, the saccharide is sized to a target molecular weight range prior to conjugation to a carrier protein.

有利地，經純化之莢膜肺炎鏈球菌血清型38醣之尺寸減小，同時保留多醣結構之關鍵特徵。可採用機械或化學尺寸設定。Advantageously, the size of the purified S. pneumoniae serotype 38 saccharide is reduced while retaining key features of the polysaccharide structure. Mechanical or chemical sizing may be employed.

在一實施例中，經純化之肺炎鏈球菌血清型38莢膜醣之尺寸藉由化學水解減小。化學水解可使用弱酸(例如乙酸、甲酸、丙酸)進行。在一實施例中，化學水解係使用甲酸進行。在一實施例中，化學水解係使用丙酸進行。在一較佳實施例中，化學水解係使用乙酸進行。化學水解亦可使用經稀釋之強酸(諸如稀鹽酸、稀硫酸、稀磷酸、稀硝酸或稀過氯酸)進行。在一實施例中，化學水解係使用稀鹽酸進行。在一實施例中，化學水解係使用烯硫酸進行。在一實施例中，化學水解係使用烯磷酸進行。在一實施例中，化學水解係使用烯硝酸進行。在一實施例中，化學水解係使用烯過氯酸進行。In one embodiment, the size of the purified S. pneumoniae serotype 38 capsular saccharide is reduced by chemical hydrolysis. Chemical hydrolysis can be performed using a weak acid (e.g., acetic acid, formic acid, propionic acid). In one embodiment, chemical hydrolysis is performed using formic acid. In one embodiment, chemical hydrolysis is performed using propionic acid. In a preferred embodiment, chemical hydrolysis is performed using acetic acid. Chemical hydrolysis can also be performed using a diluted strong acid (e.g., dilute hydrochloric acid, dilute sulfuric acid, dilute phosphoric acid, dilute nitric acid, or dilute perchloric acid). In one embodiment, chemical hydrolysis is performed using dilute hydrochloric acid. In one embodiment, chemical hydrolysis is performed using olefinic sulfuric acid. In one embodiment, chemical hydrolysis is performed using olefinic phosphoric acid. In one embodiment, chemical hydrolysis is performed using olefinic nitric acid. In one embodiment, the chemical hydrolysis is performed using perchloric acid.

經純化之肺炎鏈球菌血清型38莢膜醣之尺寸亦可藉由機械均質化減小。在一實施例中，經純化之莢膜醣之尺寸藉由高壓均質化減小。高壓均質化藉由通過具有足夠小尺寸之流徑泵送製程流體來達成高剪切速率。剪切速率藉由使用較大的所施加均質化壓力而增加，且暴露時間可藉由使進料流再循環通過均質器而增加。The size of purified S. pneumoniae serotype 38 capsular saccharides can also be reduced by mechanical homogenization. In one embodiment, the size of purified capsular saccharides is reduced by high pressure homogenization. High pressure homogenization achieves high shear rates by pumping the process fluid through a flow path with a sufficiently small size. The shear rate is increased by using a greater applied homogenization pressure, and the exposure time can be increased by recirculating the feed stream through the homogenizer.

高壓均質化製程可適用於減小經純化之莢膜醣之尺寸，同時保留醣之結構特徵。High pressure homogenization processes can be used to reduce the size of purified capsular saccharides while retaining the structural characteristics of the saccharides.

在一較佳實施例中，經分離之肺炎鏈球菌血清型38莢膜醣之尺寸設定為重量平均分子量在10 kDa與1000 kDa之間。在一實施例中，經分離之肺炎鏈球菌血清型38莢膜醣之尺寸設定為重量平均分子量在50 kDa與500 kDa之間。在一實施例中，經分離之肺炎鏈球菌血清型38莢膜醣之尺寸設定為重量平均分子量在50 kDa與400 kDa之間。在一實施例中，經分離之肺炎鏈球菌血清型38莢膜醣之尺寸設定為重量平均分子量在50 kDa與250 kDa之間。In a preferred embodiment, the size of the isolated pneumococcal serotype 38 capsaccharide is set to a weight average molecular weight between 10 kDa and 1000 kDa. In one embodiment, the size of the isolated pneumococcal serotype 38 capsaccharide is set to a weight average molecular weight between 50 kDa and 500 kDa. In one embodiment, the size of the isolated pneumococcal serotype 38 capsaccharide is set to a weight average molecular weight between 50 kDa and 400 kDa. In one embodiment, the size of the isolated pneumococcal serotype 38 capsaccharide is set to a weight average molecular weight between 50 kDa and 250 kDa.

在一較佳實施例中，經分離之肺炎鏈球菌血清型38莢膜醣之尺寸設定為重量平均分子量在100 kDa與1000 kDa之間。在一實施例中，經分離之肺炎鏈球菌血清型38莢膜醣之尺寸設定為重量平均分子量在100 kDa與500 kDa之間。在一實施例中，經分離之肺炎鏈球菌血清型38莢膜醣之尺寸設定為重量平均分子量在100 kDa與400 kDa之間。在一實施例中，經分離之肺炎鏈球菌血清型38莢膜醣之尺寸設定為重量平均分子量在100 kDa與250 kDa之間。In a preferred embodiment, the size of the isolated pneumococcal serotype 38 capsaccharide is set to a weight average molecular weight between 100 kDa and 1000 kDa. In one embodiment, the size of the isolated pneumococcal serotype 38 capsaccharide is set to a weight average molecular weight between 100 kDa and 500 kDa. In one embodiment, the size of the isolated pneumococcal serotype 38 capsaccharide is set to a weight average molecular weight between 100 kDa and 400 kDa. In one embodiment, the size of the isolated pneumococcal serotype 38 capsaccharide is set to a weight average molecular weight between 100 kDa and 250 kDa.

在一實施例中，經分離之肺炎鏈球菌血清型38莢膜醣之尺寸設定為重量平均分子量在250 kDa與1000 kDa之間。在一實施例中，經分離之肺炎鏈球菌血清型38莢膜醣之尺寸設定為重量平均分子量在250 kDa與500 kDa之間。在一實施例中，經分離之肺炎鏈球菌血清型38莢膜醣之尺寸設定為重量平均分子量在250 kDa與400 kDa之間。在一實施例中，經分離之肺炎鏈球菌血清型38莢膜醣之尺寸設定為重量平均分子量在200 kDa與800 kDa之間。In one embodiment, the size of the isolated pneumococcal serotype 38 capsaccharide is set to a weight average molecular weight between 250 kDa and 1000 kDa. In one embodiment, the size of the isolated pneumococcal serotype 38 capsaccharide is set to a weight average molecular weight between 250 kDa and 500 kDa. In one embodiment, the size of the isolated pneumococcal serotype 38 capsaccharide is set to a weight average molecular weight between 250 kDa and 400 kDa. In one embodiment, the size of the isolated pneumococcal serotype 38 capsaccharide is set to a weight average molecular weight between 200 kDa and 800 kDa.

在一較佳實施例中，經分離之肺炎鏈球菌血清型38莢膜醣之尺寸設定為重量平均分子量在150 kDa與300 kDa之間。In a preferred embodiment, the size of the isolated S. pneumoniae serotype 38 capsular saccharide is set to have a weight average molecular weight between 150 kDa and 300 kDa.

在一實施例中，經分離之肺炎鏈球菌血清型38莢膜醣之尺寸設定為重量平均分子量為約250 kDa。在一實施例中，經分離之肺炎鏈球菌血清型38莢膜醣之尺寸設定為重量平均分子量為約300 kDa。在一實施例中，經分離之肺炎鏈球菌血清型38莢膜醣之尺寸設定為重量平均分子量為約350 kDa。在一實施例中，經分離之肺炎鏈球菌血清型38莢膜醣之尺寸設定為重量平均分子量為約400 kDa。在一實施例中，經分離之肺炎鏈球菌血清型38莢膜醣之尺寸設定為重量平均分子量為約450 kDa。在一實施例中，經分離之肺炎鏈球菌血清型38莢膜醣之尺寸設定為重量平均分子量為約500 kDa。在一實施例中，經分離之肺炎鏈球菌血清型38莢膜醣之尺寸設定為重量平均分子量為約550 kDa。在一實施例中，經分離之肺炎鏈球菌血清型38莢膜醣之尺寸設定為重量平均分子量為約600 kDa。在一實施例中，經分離之肺炎鏈球菌血清型38莢膜醣之尺寸設定為重量平均分子量為約700 kDa。在一實施例中，經分離之肺炎鏈球菌血清型38莢膜醣之尺寸設定為重量平均分子量為約800 kDa。在一實施例中，經分離之肺炎鏈球菌血清型38莢膜醣之尺寸設定為重量平均分子量為約900 kDa。在一實施例中，經分離之肺炎鏈球菌血清型38莢膜醣之尺寸設定為重量平均分子量為約1000 kDa。In one embodiment, the size of the isolated pneumococcal serotype 38 capsaccharide is set to a weight average molecular weight of about 250 kDa. In one embodiment, the size of the isolated pneumococcal serotype 38 capsaccharide is set to a weight average molecular weight of about 300 kDa. In one embodiment, the size of the isolated pneumococcal serotype 38 capsaccharide is set to a weight average molecular weight of about 350 kDa. In one embodiment, the size of the isolated pneumococcal serotype 38 capsaccharide is set to a weight average molecular weight of about 400 kDa. In one embodiment, the size of the isolated pneumococcal serotype 38 capsaccharide is set to a weight average molecular weight of about 450 kDa. In one embodiment, the size of the isolated pneumococcal serotype 38 capsaccharide is set to a weight average molecular weight of about 500 kDa. In one embodiment, the size of the isolated pneumococcal serotype 38 capsaccharide is set to a weight average molecular weight of about 550 kDa. In one embodiment, the size of the isolated pneumococcal serotype 38 capsaccharide is set to a weight average molecular weight of about 600 kDa. In one embodiment, the size of the isolated pneumococcal serotype 38 capsaccharide is set to a weight average molecular weight of about 700 kDa. In one embodiment, the size of the isolated pneumococcal serotype 38 capsaccharide is set to a weight average molecular weight of about 800 kDa. In one embodiment, the size of the isolated S. pneumoniae serotype 38 capsaccharide is set to a weight average molecular weight of about 900 kDa. In one embodiment, the size of the isolated S. pneumoniae serotype 38 capsaccharide is set to a weight average molecular weight of about 1000 kDa.

在一實施例中，經分離之莢膜醣未經尺寸設定。In one embodiment, the isolated capsular saccharide has not been sized.

上文所描述之經分離之莢膜醣可經活化(例如經化學活化)，使得其能夠反應且隨後併入醣結合物中，如本文進一步所描述。The isolated capsular saccharides described above may be activated (e.g., chemically activated) so that they are able to react and subsequently be incorporated into glycoconjugates, as further described herein.

2. 本發明之肺炎鏈球菌血清型 38 醣結合物出於本發明之目的，術語『醣結合物』指示經由共價或非共價鍵與載體蛋白結合之莢膜醣。在一實施例中，莢膜醣經由非共價鍵(諸如，根瘤菌抗生物素蛋白(rhizavidin)/生物素系統，參見例如WO2012155007、WO2020056202)與載體蛋白結合。較佳地，莢膜醣經由共價鍵結合。在一個實施例中，莢膜醣與載體蛋白直接結合。在第二實施例中，莢膜醣透過間隔子/連接子與載體蛋白結合。 2. The Streptococcus pneumoniae serotype 38 glycoconjugate of the present invention For the purpose of the present invention, the term "glycoconjugate" refers to a capsular saccharide that is bound to a carrier protein via a covalent or non-covalent bond. In one embodiment, the capsular saccharide is bound to a carrier protein via a non-covalent bond (e.g., rhizavidin/biotin system, see, e.g., WO2012155007, WO2020056202). Preferably, the capsular saccharide is bound via a covalent bond. In one embodiment, the capsular saccharide is directly bound to the carrier protein. In a second embodiment, the capsular saccharide is bound to the carrier protein via a spacer/linker.

本發明提供醣結合物，其中如上文所提供之醣與載體蛋白結合。因此，在一實施例中，本發明提供一種醣結合物，其包含與載體蛋白結合的具有上文所揭露之重複單元的醣。The present invention provides a carbohydrate conjugate, wherein the carbohydrate provided above is conjugated to a carrier protein. Therefore, in one embodiment, the present invention provides a carbohydrate conjugate, which comprises a carbohydrate having the repeating unit disclosed above conjugated to a carrier protein.

在一實施例中，本發明提供一種醣結合物，其由與載體蛋白結合的具有以上所揭露之重複單元的醣組成。In one embodiment, the present invention provides a carbohydrate conjugate, which is composed of a carbohydrate having the repeating units disclosed above conjugated to a carrier protein.

2.1 本發明之肺炎鏈球菌血清型 38 醣結合物之屬性上文所描述之經分離之多醣可經活化(例如經化學活化)，使得其能夠反應(例如與連接子或直接與載體蛋白反應)且隨後併入醣結合物中，如本文進一步所描述。 2.1 Properties of the S. pneumoniae serotype 38 glycoconjugates of the present invention The isolated polysaccharides described above may be activated (eg chemically activated) so that they can react (eg with a linker or directly with a carrier protein) and subsequently be incorporated into a glycoconjugate, as further described herein.

在活化之前，經分離之多醣之尺寸可減小，同時保留多醣結構之關鍵特徵。可採用機械或化學尺寸設定。在一實施例中，經分離之多醣之尺寸藉由化學水解減小。經分離之多醣之尺寸亦可藉由機械均質化減小。在一實施例中，經分離之多醣之尺寸藉由高壓均質化減小。高壓均質化藉由通過具有足夠小尺寸之流徑泵送製程流體來達成高剪切速率。剪切速率藉由使用較大的所施加均質化壓力而增加，且暴露時間可藉由使進料流再循環通過均質器而增加。Prior to activation, the size of the separated polysaccharides can be reduced while retaining key features of the polysaccharide structure. Mechanical or chemical sizing can be used. In one embodiment, the size of the separated polysaccharides is reduced by chemical hydrolysis. The size of the separated polysaccharides can also be reduced by mechanical homogenization. In one embodiment, the size of the separated polysaccharides is reduced by high pressure homogenization. High pressure homogenization achieves high shear rates by pumping the process fluid through a flow path with a sufficiently small size. The shear rate is increased by using a larger applied homogenization pressure, and the exposure time can be increased by recirculating the feed stream through the homogenizer.

在結合之前醣之重量平均分子量(Mw)係指醣活化之前(亦即在最終尺寸設定步驟之後但在醣與活化劑反應之前)的Mw。在本發明之上下文中，醣之Mw未由活化步驟實質上改變，且併入結合物中的醣之Mw與活化之前量測的醣之Mw類似。The weight average molecular weight (Mw) of the carbohydrate prior to conjugation refers to the Mw of the carbohydrate prior to activation (i.e., after the final sizing step but before the carbohydrate is reacted with the activating agent). In the context of the present invention, the Mw of the carbohydrate is not substantially altered by the activation step, and the Mw of the carbohydrate incorporated into the conjugate is similar to the Mw of the carbohydrate measured prior to activation.

在一實施例中，本發明之血清型38醣結合物包含血清型38多醣，其中在結合之前該多醣之重量平均分子量(Mw)在50 kDa與1,000 kDa之間。In one embodiment, the serotype 38 carbohydrate conjugate of the present invention comprises a serotype 38 polysaccharide, wherein the weight average molecular weight (Mw) of the polysaccharide before conjugation is between 50 kDa and 1,000 kDa.

在一實施例中，本發明之血清型38醣結合物包含血清型38多醣，其中在結合之前該多醣之重量平均分子量(Mw)在100 kDa與600 kDa之間。In one embodiment, the serotype 38 carbohydrate conjugate of the present invention comprises serotype 38 polysaccharide, wherein the weight average molecular weight (Mw) of the polysaccharide before conjugation is between 100 kDa and 600 kDa.

在一實施例中，本發明之血清型38醣結合物包含血清型38多醣，其中在結合之前該多醣之重量平均分子量(Mw)在100 kDa與400 kDa之間。In one embodiment, the serotype 38 carbohydrate conjugate of the present invention comprises serotype 38 polysaccharide, wherein the weight average molecular weight (Mw) of the polysaccharide before conjugation is between 100 kDa and 400 kDa.

在一較佳實施例中，本發明之血清型38醣結合物包含血清型38多醣，其中在結合之前該多醣之重量平均分子量(Mw)在150 kDa與300 kDa之間。In a preferred embodiment, the serotype 38 carbohydrate conjugate of the present invention comprises serotype 38 polysaccharide, wherein the weight average molecular weight (Mw) of the polysaccharide before conjugation is between 150 kDa and 300 kDa.

在一些實施例中，本發明之血清型38醣結合物之重量平均分子量(Mw)在250 kDa 與20,000 kDa之間。In some embodiments, the weight average molecular weight (Mw) of the serotype 38 glycoconjugate of the present invention is between 250 kDa and 20,000 kDa.

在其他實施例中，血清型38醣結合物之重量平均分子量(Mw)在500 kDa與15,000 kDa之間。In other embodiments, the weight average molecular weight (Mw) of the serotype 38 glycoconjugate is between 500 kDa and 15,000 kDa.

在其他實施例中，血清型38醣結合物之重量平均分子量(Mw)在500 kDa與10,000 kDa之間。In other embodiments, the weight average molecular weight (Mw) of the serotype 38 glycoconjugate is between 500 kDa and 10,000 kDa.

在又其他實施例中，血清型38醣結合物之重量平均分子量(Mw)在750 kDa與7,500 kDa之間。In yet other embodiments, the weight average molecular weight (Mw) of the serotype 38 glycoconjugate is between 750 kDa and 7,500 kDa.

在其他實施例中，血清型38醣結合物之重量平均分子量(Mw)在1,000 kDa與5,000 kDa 之間。In other embodiments, the weight average molecular weight (Mw) of the serotype 38 glycoconjugate is between 1,000 kDa and 5,000 kDa.

在較佳實施例中，血清型38醣結合物之重量平均分子量(Mw)在1,000 kDa與10,000 kDa之間。In a preferred embodiment, the weight average molecular weight (Mw) of the serotype 38 glycoconjugate is between 1,000 kDa and 10,000 kDa.

另一表徵本發明之血清型38醣結合物的方式為載體蛋白中之離胺酸殘基(例如，CRM 197、SCP、DT或TT)的數目，其與醣結合，該醣可表徵為經結合離胺酸之範圍(結合程度)。可藉由胺基酸分析且使用熟習此項技術者已知之常規方法來獲得載體蛋白之離胺酸修飾(歸因於與多醣之共價連接)之證據。與用於產生結合物物質之載體蛋白起始物質相比，結合引起所回收之離胺酸殘基之數目減少。在一較佳實施例中，本發明之血清型38醣結合物的結合程度在2與15之間。在一實施例中，本發明之血清型38醣結合物的結合程度在2與10之間。在一實施例中，本發明之血清型38醣結合物的結合程度在3與5之間。在一實施例中，本發明之血清型38醣結合物的結合程度在2與6之間。在一較佳實施例中，本發明之血清型38醣結合物的結合程度在4與10之間。 Another way to characterize the serotype 38 glycoconjugates of the invention is the number of lysine residues (e.g., CRM 197 , SCP, DT or TT) in the carrier protein that are conjugated to a sugar that can be characterized as the extent of conjugated lysine (degree of conjugation). Evidence of lysine modification of the carrier protein (due to covalent attachment to the polysaccharide) can be obtained by amino acid analysis using conventional methods known to those skilled in the art. Conjugation results in a reduction in the number of lysine residues recovered compared to the carrier protein starting material used to generate the conjugate material. In a preferred embodiment, the degree of conjugation of the serotype 38 glycoconjugates of the invention is between 2 and 15. In one embodiment, the serotype 38 saccharide conjugate of the present invention has a binding degree between 2 and 10. In one embodiment, the serotype 38 saccharide conjugate of the present invention has a binding degree between 3 and 5. In one embodiment, the serotype 38 saccharide conjugate of the present invention has a binding degree between 2 and 6. In a preferred embodiment, the serotype 38 saccharide conjugate of the present invention has a binding degree between 4 and 10.

本發明之血清型38醣結合物亦可藉由醣與載體蛋白之比率(重量/重量)表徵。在一些實施例中，醣結合物中血清型38醣與載體蛋白之比率(w/w)在0.5與3.0之間。在其他實施例中，醣與載體蛋白之比率(w/w)在0.5與2.0之間。在其他實施例中，醣與載體蛋白之比率(w/w)在0.5與1.5之間。在其他實施例中，醣與載體蛋白之比率(w/w)在0.8與1.2之間。在其他實施例中，醣與載體蛋白之比率(w/w)在0.5與1.0之間。在其他實施例中，醣與載體蛋白之比率(w/w)在1.0與1.5之間。在其他實施例中，醣與載體蛋白之比率(w/w)在1.0與2.0之間。在其他實施例中，醣與載體蛋白之比率(w/w)在0.8與1.2之間。在其他實施例中，結合物中血清型38醣與載體蛋白之比率在0.7與1.1之間。在較佳實施例中，結合物中血清型38醣與載體蛋白之比率在0.5與1.5之間。在一些此類實施例中，載體蛋白質為CRM 197。在一些較佳實施例中，載體蛋白為SCP。 The serotype 38 glycoconjugates of the present invention can also be characterized by the ratio of serotype 38 serotype 38 serotype 38 glycoconjugates to carrier proteins (w/w). In some embodiments, the ratio of serotype 38 serotype 38 serotype 38 glycoconjugates to carrier proteins (w/w) is between 0.5 and 3.0. In other embodiments, the ratio of serotype 38 serotype 38 glycoconjugates to carrier proteins (w/w) is between 0.5 and 2.0. In other embodiments, the ratio of serotype 38 serotype 38 glycoconjugates to carrier proteins (w/w) is between 0.5 and 1.5. In other embodiments, the ratio of serotype 38 serotype 38 glycoconjugates to carrier proteins (w/w) is between 0.8 and 1.2. In other embodiments, the ratio of serotype 38 serotype 38 glycoconjugates to carrier proteins (w/w) is between 0.5 and 1.0. In other embodiments, the ratio of serotype 38 serotype 38 glycoconjugates to carrier proteins (w/w) is between 1.0 and 1.5. In other embodiments, the ratio of carbohydrate to carrier protein (w/w) is between 1.0 and 2.0. In other embodiments, the ratio of carbohydrate to carrier protein (w/w) is between 0.8 and 1.2. In other embodiments, the ratio of serotype 38 carbohydrate to carrier protein in the conjugate is between 0.7 and 1.1. In preferred embodiments, the ratio of serotype 38 carbohydrate to carrier protein in the conjugate is between 0.5 and 1.5. In some such embodiments, the carrier protein is CRM 197 . In some preferred embodiments, the carrier protein is SCP.

本發明之血清型38醣結合物及免疫原組合物可含有未與載體蛋白結合但仍存在於醣結合物組合物中之游離醣。游離醣可與醣結合物非共價締合(亦即，與醣結合物非共價鍵結、吸附至醣結合物、或者包覆於醣結合物中或由醣結合物包覆)。在一較佳實施例中，血清型38醣結合物包含與血清型38醣之總量相比小於約50%之游離血清型38醣。在一較佳實施例中，血清型38醣結合物包含與血清型38醣之總量相比小於約25%之游離血清型38醣。在一甚至較佳實施例中，血清型38醣結合物包含與血清型38醣之總量相比小於約20%之游離血清型38醣。在又一較佳實施例中，血清型38醣結合物包含與血清型38醣之總量相比小於約15%之游離血清型38醣。The serotype 38 saccharide conjugates and immunogenic compositions of the present invention may contain free saccharides that are not bound to a carrier protein but are still present in the saccharide conjugate composition. The free saccharides may be non-covalently associated with the saccharide conjugate (i.e., non-covalently bonded to, adsorbed to, or encapsulated in or by the saccharide conjugate). In a preferred embodiment, the serotype 38 saccharide conjugate comprises less than about 50% free serotype 38 saccharides compared to the total amount of serotype 38 saccharides. In a preferred embodiment, the serotype 38 saccharide conjugate comprises less than about 25% free serotype 38 saccharides compared to the total amount of serotype 38 saccharides. In an even more preferred embodiment, the serotype 38 carbohydrate conjugate contains less than about 20% free serotype 38 carbohydrate compared to the total amount of serotype 38 carbohydrate. In another preferred embodiment, the serotype 38 carbohydrate conjugate contains less than about 15% free serotype 38 carbohydrate compared to the total amount of serotype 38 carbohydrate.

血清型38醣結合物亦可藉由其分子尺寸分佈(K d)表徵。尺寸排阻層析介質(CL-4B)可用於測定結合物之相對分子尺寸分佈。在重力饋送柱中使用尺寸排阻層析(SEC)以得到結合物之分子尺寸分佈概況。自介質之孔中排阻之大分子比小分子溶離得更快。使用溶離份收集器來收集管柱溶離液。藉由醣分析法來對溶離份進行比色測試。為了測定K d，將管柱進行校準，以確立分子被完全排阻之分數(V 0), (K d=0)及表示最大保留之分數(V i), (K d=1)。達到指定樣品屬性之分數(V e)藉由表達式K d= (V e- V 0)/(V i- V 0)與K d相關。 Serotype 38 glycoconjugates can also be characterized by their molecular size distribution ( Kd ). Size exclusion chromatography medium (CL-4B) can be used to determine the relative molecular size distribution of the conjugate. Size exclusion chromatography (SEC) is used in a gravity fed column to obtain a molecular size distribution profile of the conjugate. Large molecules excluded from the pores of the medium elute faster than small molecules. A fraction collector is used to collect the column eluate. The fractions are assayed colorimetrically by glycoanalysis. To determine Kd , the column is calibrated to determine the fraction ( V0 ) at which the molecule is completely excluded, ( Kd = 0) and the fraction ( Vi ) representing maximal retention, ( Kd = 1). The fraction (V e ) that achieves a given sample property is related to K d by the expression K d = (V e - V 0 )/(V i - V 0 ).

在一較佳實施例中，至少30%的該血清型38醣結合物在CL-4B管柱中之K d低於或等於0.3。在一較佳實施例中，至少40%的該醣結合物在CL-4B管柱中之K d低於或等於0.3。在一較佳實施例中，至少60%的該血清型38醣結合物在CL-4B管柱中之K d低於或等於0.3。在一較佳實施例中，50%與80%之間的該血清型38醣結合物在CL-4B管柱中之K d低於或等於0.3。在一較佳實施例中，65%與80%之間的該血清型38醣結合物在CL-4B管柱中之K d低於或等於0.3。 In a preferred embodiment, at least 30% of the serotype 38 glycoconjugates have a K d of less than or equal to 0.3 in a CL-4B column. In a preferred embodiment, at least 40% of the serotype 38 glycoconjugates have a K d of less than or equal to 0.3 in a CL-4B column. In a preferred embodiment, at least 60% of the serotype 38 glycoconjugates have a K d of less than or equal to 0.3 in a CL-4B column. In a preferred embodiment, between 50% and 80% of the serotype 38 glycoconjugates have a K d of less than or equal to 0.3 in a CL-4B column. In a preferred embodiment, between 65% and 80% of the serotype 38 glycoconjugates have a K d of less than or equal to 0.3 in a CL-4B column.

2.2 帶有經還原之 α - D - 2 - 乙醯胺基 - 2 , 6 - 二去氧基 - 木糖 - 己 - 4 - 酮糖 ( α - D - Sug ) 殘基之本發明之肺炎鏈球菌血清型 38 醣結合物製備本發明之血清型38醣結合物之方法可包含使用還原劑。特定言之，可使用適合的封端劑(還原劑)封端氧化後(尤其當使用還原胺化時，參見下文)的未反應之醛基。在一個實施例中，此封端劑為硼氫化鈉(NaBH 4)。 2.2 Streptococcus pneumoniae serotype 38 glycoconjugates of the present invention with reduced α - D - 2 - acetamido - 2,6 - dideoxy - xylose - hex - 4 - ketose ( α - D - Sug ) residues The method for preparing the serotype 38 glycoconjugates of the present invention may comprise the use of a reducing agent. In particular, a suitable capping agent ( reducing agent) may be used to cap unreacted aldehyde groups after oxidation (especially when reductive amination is used, see below). In one embodiment, the capping agent is sodium borohydride (NaBH 4 ).

如實例4所示，α-D-2-乙醯胺基-2,6-二去氧基-木糖-己-4-酮糖(α-D-Sug)殘基對使用NaBH 4還原敏感。用NaBH 4處理血清型38多醣特異性地將D-Sug殘基之位置4自酮/水合物還原成醇，且將D-Sug轉化成D-FucNAc及D-QuiNAc之混合物，其特徵為位置4羥基分別處於軸向及赤道取向，如圖6中所示。 As shown in Example 4, the α-D-2-acetamido-2,6-dideoxy-xylose-hex-4-ketose (α-D-Sug) residue is sensitive to reduction using NaBH 4. Treatment of serotype 38 polysaccharide with NaBH 4 specifically reduces position 4 of the D-Sug residue from the keto/hydrate to the alcohol and converts D-Sug to a mixture of D-FucNAc and D-QuiNAc, which are characterized by the hydroxyl group at position 4 being in the axial and equatorial orientations, respectively, as shown in FIG6 .

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約1至約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 1 to about 70 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約1至約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 1 to about 68 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約1至約65個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 1 to about 65 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約1至約60個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 1 to about 60 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約1至約50個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 1 to about 50 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約1至約40個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 1 to about 40 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約1至約30個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 1 to about 30 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約1至約20個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 1 to about 20 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約1至約10個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 1 to about 10 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約1至約5個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 1 to about 5 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約10至約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 10 to about 70 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約10至約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 10 to about 68 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約10至約65個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 10 to about 65 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約10至約60個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 10 to about 60 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約10至約50個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 10 to about 50 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約10至約40個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 10 to about 40 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約10至約30個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 10 to about 30 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約10至約20個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 10 to about 20 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約20至約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 20 to about 70 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約20至約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 20 to about 68 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約20至約65個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 20 to about 65 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約20至約60個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 20 to about 60 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約20至約50個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 20 to about 50 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約20至約40個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 20 to about 40 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約20至約30個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 20 to about 30 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約40至約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 40 to about 70 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約40至約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 40 to about 68 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約40至約65個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 40 to about 65 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約40至約60個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 40 to about 60 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約40至約50個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 40 to about 50 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約50至約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 50 to about 70 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約50至約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 50 to about 68 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約50至約65個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 50 to about 65 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約50至約60個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 50 to about 60 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約60至約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 60 to about 70 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約60至約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 60 to about 68 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約60至約65個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 60 to about 65 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide comprising about 70 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide comprising about 68 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約65個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide comprising about 65 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約60個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide comprising about 60 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約50個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide comprising about 50 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約40個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide comprising about 40 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約30個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide comprising about 30 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約20個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide comprising about 20 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約10個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide comprising about 10 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約5個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide comprising about 5 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約1個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 cancellous saccharide comprising about 1 N-acetyl-D-fucosamine (D-FucNAc) residue per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約0.5至約35個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 0.5 to about 35 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約0.5至約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 0.5 to about 32 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約0.5至約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 0.5 to about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約0.5至約25個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 0.5 to about 25 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約0.5至約20個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 0.5 to about 20 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約0.5至約15個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 0.5 to about 15 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約0.5至約10個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 0.5 to about 10 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約0.5至約5個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 0.5 to about 5 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約0.5至約2.5個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 0.5 to about 2.5 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約5至約35個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 5 to about 35 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約5至約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 5 to about 32 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約5至約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 5 to about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約5至約25個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 5 to about 25 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約5至約20個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 5 to about 20 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約5至約15個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 5 to about 15 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約5至約10個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 5 to about 10 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約10至約35個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 10 to about 35 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約10至約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 10 to about 32 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約10至約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 10 to about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約10至約25個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 10 to about 25 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約10至約20個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 10 to about 20 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約10至約15個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 10 to about 15 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約20至約35個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 20 to about 35 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約20至約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 20 to about 32 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約20至約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 20 to about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約20至約25個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 20 to about 25 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約25至約35個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 25 to about 35 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約25至約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 25 to about 32 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約25至約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 25 to about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約30至約35個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 30 to about 35 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約30至約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 30 to about 32 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約35個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 35 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 canangiocarboxylic acid saccharide comprising about 32 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 canangiocarboxylic acid saccharide comprising about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約25個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 25 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約20個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 20 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約15個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 canangiocarboxylic acid saccharide comprising about 15 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約10個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 canangiocarboxylic acid saccharide comprising about 10 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約5個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 5 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約2.5個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 canangiocarboxylic acid saccharide comprising about 2.5 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約0.5個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 canangiocarboxylic acid saccharide comprising about 0.5 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約1至約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約0.5至約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 1 to about 70 N-acetyl-D-fucosamine (D-FucNAc) residues and about 0.5 to about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約1至約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約0.5至約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 1 to about 68 N-acetyl-D-fucosamine (D-FucNAc) residues and about 0.5 to about 32 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約1至約65個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約0.5至約32.5個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 1 to about 65 N-acetyl-D-fucosamine (D-FucNAc) residues and about 0.5 to about 32.5 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約1至約60個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約0.5至約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 1 to about 60 N-acetyl-D-fucosamine (D-FucNAc) residues and about 0.5 to about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約1至約50個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約0.5至約25個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 1 to about 50 N-acetyl-D-fucosamine (D-FucNAc) residues and about 0.5 to about 25 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約1至約40個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約0.5至約20個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 1 to about 40 N-acetyl-D-fucosamine (D-FucNAc) residues and about 0.5 to about 20 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約1至約30個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約0.5至約15個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 1 to about 30 N-acetyl-D-fucosamine (D-FucNAc) residues and about 0.5 to about 15 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約1至約20個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約0.5至約10個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 1 to about 20 N-acetyl-D-fucosamine (D-FucNAc) residues and about 0.5 to about 10 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約1至約10個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約0.5至約5個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 1 to about 10 N-acetyl-D-fucosamine (D-FucNAc) residues and about 0.5 to about 5 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約1至約5個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約0.5至約2.5個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 1 to about 5 N-acetyl-D-fucosamine (D-FucNAc) residues and about 0.5 to about 2.5 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約1至約2個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約0.5至約1個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 1 to about 2 N-acetyl-D-fucosamine (D-FucNAc) residues and about 0.5 to about 1 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約5至約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約2.5至約35個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 5 to about 70 N-acetyl-D-fucosamine (D-FucNAc) residues and about 2.5 to about 35 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約5至約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約2.5至約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 5 to about 68 N-acetyl-D-fucosamine (D-FucNAc) residues and about 2.5 to about 32 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約5至約65個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約2.5至約32.5個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 5 to about 65 N-acetyl-D-fucosamine (D-FucNAc) residues and about 2.5 to about 32.5 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約5至約60個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約2.5至約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 5 to about 60 N-acetyl-D-fucosamine (D-FucNAc) residues and about 2.5 to about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約5至約50個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約2.5至約25個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 5 to about 50 N-acetyl-D-fucosamine (D-FucNAc) residues and about 2.5 to about 25 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約5至約40個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約2.5至約20個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 5 to about 40 N-acetyl-D-fucosamine (D-FucNAc) residues and about 2.5 to about 20 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約5至約30個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約2.5至約15個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 5 to about 30 N-acetyl-D-fucosamine (D-FucNAc) residues and about 2.5 to about 15 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約5至約20個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約2.5至約10個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 5 to about 20 N-acetyl-D-fucosamine (D-FucNAc) residues and about 2.5 to about 10 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約5至約10個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約2.5至約5個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 5 to about 10 N-acetyl-D-fucosamine (D-FucNAc) residues and about 2.5 to about 5 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約10至約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約5至約35個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 10 to about 70 N-acetyl-D-fucosamine (D-FucNAc) residues and about 5 to about 35 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約10至約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約5至約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 10 to about 68 N-acetyl-D-fucosamine (D-FucNAc) residues and about 5 to about 32 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約10至約65個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約5至約32.5個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 10 to about 65 N-acetyl-D-fucosamine (D-FucNAc) residues and about 5 to about 32.5 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約10至約60個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約5至約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 10 to about 60 N-acetyl-D-fucosamine (D-FucNAc) residues and about 5 to about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約10至約50個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約5至約25個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 10 to about 50 N-acetyl-D-fucosamine (D-FucNAc) residues and about 5 to about 25 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約10至約40個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約5至約20個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 10 to about 40 N-acetyl-D-fucosamine (D-FucNAc) residues and about 5 to about 20 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約10至約30個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約5至約15個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 10 to about 30 N-acetyl-D-fucosamine (D-FucNAc) residues and about 5 to about 15 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約10至約20個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約5至約10個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 10 to about 20 N-acetyl-D-fucosamine (D-FucNAc) residues and about 5 to about 10 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約20至約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約10至約35個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 20 to about 70 N-acetyl-D-fucosamine (D-FucNAc) residues and about 10 to about 35 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約20至約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約10至約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 20 to about 68 N-acetyl-D-fucosamine (D-FucNAc) residues and about 10 to about 32 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約20至約65個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約10至約32.5個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 20 to about 65 N-acetyl-D-fucosamine (D-FucNAc) residues and about 10 to about 32.5 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約20至約60個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約10至約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 20 to about 60 N-acetyl-D-fucosamine (D-FucNAc) residues and about 10 to about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約20至約50個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約10至約25個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 20 to about 50 N-acetyl-D-fucosamine (D-FucNAc) residues and about 10 to about 25 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約20至約40個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約10至約20個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 20 to about 40 N-acetyl-D-fucosamine (D-FucNAc) residues and about 10 to about 20 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約20至約30個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約10至約15個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 20 to about 30 N-acetyl-D-fucosamine (D-FucNAc) residues and about 10 to about 15 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約30至約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約15至約35個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 30 to about 70 N-acetyl-D-fucosamine (D-FucNAc) residues and about 15 to about 35 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約30至約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約15至約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 30 to about 68 N-acetyl-D-fucosamine (D-FucNAc) residues and about 15 to about 32 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約30至約65個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約15至約32.5個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 30 to about 65 N-acetyl-D-fucosamine (D-FucNAc) residues and about 15 to about 32.5 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約30至約60個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約15至約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 30 to about 60 N-acetyl-D-fucosamine (D-FucNAc) residues and about 15 to about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約30至約50個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約15至約25個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 30 to about 50 N-acetyl-D-fucosamine (D-FucNAc) residues and about 15 to about 25 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約30至約40個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約15至約20個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 30 to about 40 N-acetyl-D-fucosamine (D-FucNAc) residues and about 15 to about 20 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約40至約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約20至約35個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 40 to about 70 N-acetyl-D-fucosamine (D-FucNAc) residues and about 20 to about 35 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約40至約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約20至約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 40 to about 68 N-acetyl-D-fucosamine (D-FucNAc) residues and about 20 to about 32 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約40至約65個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約20至約32.5個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 40 to about 65 N-acetyl-D-fucosamine (D-FucNAc) residues and about 20 to about 32.5 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約40至約60個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約20至約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 40 to about 60 N-acetyl-D-fucosamine (D-FucNAc) residues and about 20 to about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約40至約50個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約20至約25個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 40 to about 50 N-acetyl-D-fucosamine (D-FucNAc) residues and about 20 to about 25 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約50至約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約25至約35個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 50 to about 70 N-acetyl-D-fucosamine (D-FucNAc) residues and about 25 to about 35 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約50至約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約25至約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 50 to about 68 N-acetyl-D-fucosamine (D-FucNAc) residues and about 25 to about 32 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約50至約65個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約25至約32.5個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 50 to about 65 N-acetyl-D-fucosamine (D-FucNAc) residues and about 25 to about 32.5 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約50至約60個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約25至約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 50 to about 60 N-acetyl-D-fucosamine (D-FucNAc) residues and about 25 to about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約60至約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約30至約35個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 60 to about 70 N-acetyl-D-fucosamine (D-FucNAc) residues and about 30 to about 35 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約60至約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約30至約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 60 to about 68 N-acetyl-D-fucosamine (D-FucNAc) residues and about 30 to about 32 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約60至約65個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約30至約32.5個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide comprising about 60 to about 65 N-acetyl-D-fucosamine (D-FucNAc) residues and about 30 to about 32.5 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約1個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約0.5個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide, which comprises about 1 N-acetyl-D-fucosamine (D-FucNAc) residue and about 0.5 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約2個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約1個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide, which comprises about 2 N-acetyl-D-fucosamine (D-FucNAc) residues and about 1 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約5個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約2.5個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide comprising about 5 N-acetyl-D-fucosamine (D-FucNAc) residues and about 2.5 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約10個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約5個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide comprising about 10 N-acetyl-D-fucosamine (D-FucNAc) residues and about 5 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約20個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約10個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide comprising about 20 N-acetyl-D-fucosamine (D-FucNAc) residues and about 10 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約30個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約15個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide comprising about 30 N-acetyl-D-fucosamine (D-FucNAc) residues and about 15 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約40個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約20個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide comprising about 40 N-acetyl-D-fucosamine (D-FucNAc) residues and about 20 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約50個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約25個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide comprising about 50 N-acetyl-D-fucosamine (D-FucNAc) residues and about 25 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約60個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide, which comprises about 60 N-acetyl-D-fucosamine (D-FucNAc) residues and about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide, which comprises about 68 N-acetyl-D-fucosamine (D-FucNAc) residues and about 32 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，其在該醣的每100個醣重複單元中包含約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約35個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide, which comprises about 70 N-acetyl-D-fucosamine (D-FucNAc) residues and about 35 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，該莢膜醣包含N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基，其中較佳地D-FucNAc殘基之數目為約D-QuiNAc殘基之數目的兩倍。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide, which comprises N-acetyl-D-fucosamine (D-FucNAc) residues and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues, wherein preferably the number of D-FucNAc residues is about twice the number of D-QuiNAc residues.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，該莢膜醣包含N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基，其中較佳地D-FucNAc殘基之數目為約D-QuiNAc殘基之數目的兩倍。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide, which comprises N-acetyl-D-fucosamine (D-FucNAc) residues and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues, wherein preferably the number of D-FucNAc residues is about twice the number of D-QuiNAc residues.

在以上實施例中之任一者中，重複單元中相同位置處的剩餘糖殘基可為D-2-乙醯胺基-2,6-二去氧基-木糖-己-4-酮糖(D-Sug)。因此，在一個實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，該莢膜醣包含N-乙醯基-D-岩藻糖胺(D-FucNAc)、N-乙醯基-D-奎諾糠胺(D-QuiNAc)及D-2-乙醯胺基-2,6-二去氧基-木糖-己-4-酮糖(D-Sug)殘基。In any of the above embodiments, the remaining sugar residue at the same position in the repeating unit may be D-2-acetamido-2,6-dideoxy-xylose-hex-4-ketose (D-Sug). Therefore, in one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 canangiocarboxylic acid, which contains N-acetyl-D-fucosamine (D-FucNAc), N-acetyl-D-quinofurylamine (D-QuiNAc) and D-2-acetamido-2,6-dideoxy-xylose-hex-4-ketose (D-Sug) residue.

在一實施例中，本發明之血清型38醣結合物包含血清型38莢膜醣，該莢膜醣在重複單元之相同位置處包含N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基、N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基或D-2-乙醯胺基-2,6-二去氧基-木糖-己-4-酮糖(D-Sug)殘基。在一實施例中，D-FucNAc殘基之數目係約D-QuiNAc殘基之數目的兩倍。In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 canangiocarboxylic acid, which contains N-acetyl-D-fucosamine (D-FucNAc) residues, N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues or D-2-acetamido-2,6-dideoxy-xylose-hex-4-ketose (D-Sug) residues at the same position of the repeating unit. In one embodiment, the number of D-FucNAc residues is about twice the number of D-QuiNAc residues.

在一實施例中，本發明之血清型38醣結合物包含具有以下重複單元之血清型38莢膜醣： 其中n表示重複單元之數目，其中Y表示絲胺酸或甘胺酸殘基，其中所有該等重複單元包含相同的Y殘基，且其中X表示N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。在一實施例中，該血清型38莢膜醣在該醣的每100個醣重複單元中包含約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。在一實施例中，該血清型38莢膜醣在該醣的每100個醣重複單元中包含約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。 In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide having the following repeating units: wherein n represents the number of repeating units, wherein Y represents a serine or glycine residue, wherein all of the repeating units comprise the same Y residue, and wherein X represents an N-acetyl-D-fucosamine (D-FucNAc) residue or an N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue. In one embodiment, the serotype 38 capsaccharide comprises about 70 N-acetyl-D-fucosamine (D-FucNAc) residues and about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeating units of the saccharide. In one embodiment, the serotype 38 capsular saccharide comprises about 68 N-acetyl-D-fucosamine (D-FucNAc) residues and about 32 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，基本上所有的D-Sug殘基均被還原。因此，在一實施例中，本發明之血清型38醣結合物包含具有以下重複單元之血清型38莢膜醣： 其中n表示重複單元之數目，其中Y表示絲胺酸或甘胺酸殘基，其中所有該等重複單元包含相同的Y殘基，且其中X表示N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。在一實施例中，該血清型38莢膜醣在該醣的每100個醣重複單元中包含約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。在一實施例中，該血清型38莢膜醣在該醣的每100個醣重複單元中包含約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。 In one embodiment, substantially all D-Sug residues are reduced. Therefore, in one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide having the following repeating units: wherein n represents the number of repeating units, wherein Y represents a serine or glycine residue, wherein all of the repeating units comprise the same Y residue, and wherein X represents an N-acetyl-D-fucosamine (D-FucNAc) residue or an N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue. In one embodiment, the serotype 38 capsaccharide comprises about 70 N-acetyl-D-fucosamine (D-FucNAc) residues and about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeating units of the saccharide. In one embodiment, the serotype 38 capsular saccharide comprises about 68 N-acetyl-D-fucosamine (D-FucNAc) residues and about 32 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含具有以下重複單元之血清型38莢膜醣： 其中n表示重複單元之數目且其中X表示N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基、N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基或D-2-乙醯胺基-2,6-二去氧基-木糖-己-4-酮糖(D-Sug)殘基。在一實施例中，該血清型38莢膜醣在該醣的每100個醣重複單元中包含約1至約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基、約0.5至約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基及約0至約98.5個D-2-乙醯胺基-2,6-二去氧基-木糖-己-4-酮糖(D-Sug)殘基。 In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide having the following repeating units: wherein n represents the number of repeating units and wherein X represents an N-acetyl-D-fucosamine (D-FucNAc) residue, an N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue or a D-2-acetamido-2,6-dideoxy-xylose-hex-4-ketose (D-Sug) residue. In one embodiment, the serotype 38 capsular saccharide comprises about 1 to about 70 N-acetyl-D-fucosamine (D-FucNAc) residues, about 0.5 to about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues, and about 0 to about 98.5 D-2-acetamido-2,6-dideoxy-xylose-hex-4-ketose (D-Sug) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，基本上所有的D-Sug殘基均被還原。因此，在一實施例中，本發明之血清型38醣結合物包含具有以下重複單元之血清型38莢膜醣： 其中n表示重複單元之數目且其中X表示N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。在一實施例中，該血清型38莢膜醣在該醣的每100個醣重複單元中包含約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。在一實施例中，該血清型38莢膜醣在該醣的每100個醣重複單元中包含約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。 In one embodiment, substantially all D-Sug residues are reduced. Therefore, in one embodiment, the serotype 38 glycoconjugate of the present invention comprises a serotype 38 capsular saccharide having the following repeating units: wherein n represents the number of repeating units and wherein X represents an N-acetyl-D-fucosamine (D-FucNAc) residue or an N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue. In one embodiment, the serotype 38 capsane contains about 70 N-acetyl-D-fucosamine (D-FucNAc) residues and about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeating units of the saccharide. In one embodiment, the serotype 38 capsular saccharide comprises about 68 N-acetyl-D-fucosamine (D-FucNAc) residues and about 32 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，本發明之血清型38醣結合物包含具有以下重複單元之血清型38莢膜醣： 其中n表示重複單元之數目且其中X表示N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基、N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基或D-2-乙醯胺基-2,6-二去氧基-木糖-己-4-酮糖(D-Sug)殘基。在一實施例中，該血清型38莢膜醣在該醣的每100個醣重複單元中包含約1至約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基、約0.5至約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基及約0至約98.5個D-2-乙醯胺基-2,6-二去氧基-木糖-己-4-酮糖(D-Sug)殘基。 In one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide having the following repeating units: wherein n represents the number of repeating units and wherein X represents an N-acetyl-D-fucosamine (D-FucNAc) residue, an N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue or a D-2-acetamido-2,6-dideoxy-xylose-hex-4-ketose (D-Sug) residue. In one embodiment, the serotype 38 capsular saccharide comprises about 1 to about 70 N-acetyl-D-fucosamine (D-FucNAc) residues, about 0.5 to about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues, and about 0 to about 98.5 D-2-acetamido-2,6-dideoxy-xylose-hex-4-ketose (D-Sug) residues per 100 carbohydrate repeat units of the saccharide.

在一實施例中，基本上所有的D-Sug殘基均被還原。因此，在一實施例中，基本上所有的D-Sug殘基均被還原。因此，在一實施例中，本發明之血清型38醣結合物包含具有以下重複單元之血清型38莢膜醣： 其中n表示重複單元之數目且其中X表示N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。在一實施例中，該血清型38莢膜醣在該醣的每100個醣重複單元中包含約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。在一實施例中，該血清型38莢膜醣在該醣的每100個醣重複單元中包含約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。 In one embodiment, substantially all D-Sug residues are reduced. Therefore, in one embodiment, substantially all D-Sug residues are reduced. Therefore, in one embodiment, the serotype 38 glycoconjugate of the present invention comprises serotype 38 capsular saccharide having the following repeating units: wherein n represents the number of repeating units and wherein X represents an N-acetyl-D-fucosamine (D-FucNAc) residue or an N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue. In one embodiment, the serotype 38 capsane contains about 70 N-acetyl-D-fucosamine (D-FucNAc) residues and about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeating units of the saccharide. In one embodiment, the serotype 38 capsular saccharide comprises about 68 N-acetyl-D-fucosamine (D-FucNAc) residues and about 32 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide.

2.3 本發明之肺炎鏈球菌血清型 38 醣結合物之製備模式本發明之血清型38醣結合物可藉由一般熟習此項技術者已知之任何偶合技術製備。 2.3 Preparation Mode of the S. pneumoniae serotype 38 Sugar Conjugate of the Present Invention The serotype 38 sugar conjugate of the present invention can be prepared by any coupling technique known to those skilled in the art.

在一實施例中，血清型38醣經由非共價鍵與載體蛋白偶合(參見例如WO2012155007、WO2020056202)。In one embodiment, the serotype 38 carbohydrate is coupled to the carrier protein via a non-covalent bond (see, for example, WO2012155007, WO2020056202).

在一實施例中，血清型38醣經由共價鍵結合。在一個實施例中，莢膜醣與載體蛋白直接結合。在第二實施例中，莢膜醣透過間隔子/連接子與載體蛋白結合。In one embodiment, the serotype 38 saccharide is bound via a covalent bond. In one embodiment, the capsular saccharide is directly bound to the carrier protein. In a second embodiment, the capsular saccharide is bound to the carrier protein via a spacer/linker.

在一實施例中，本發明之血清型38醣結合物經由連接子(例如雙官能連接子)與載體蛋白結合。連接子視情況為異雙官能或同雙官能，具有例如一個反應性胺基及一個反應性羧酸基、兩個反應性胺基或兩個反應性羧酸基。連接子具有例如4與20個、4與12個、5與10個之間的碳原子。In one embodiment, the serotype 38 glycoconjugate of the present invention is conjugated to a carrier protein via a linker (e.g., a bifunctional linker). The linker is optionally heterobifunctional or homobifunctional, having, for example, one reactive amine group and one reactive carboxylic acid group, two reactive amine groups, or two reactive carboxylic acid groups. The linker has, for example, between 4 and 20, 4 and 12, 5 and 10 carbon atoms.

可能的連接子為己二酸二醯肼(ADH)。其他連接子包括B-丙醯胺基(WO 00/10599)、硝基苯基-乙胺(Gever等人(1979) Med. Microbiol. lmmunol. 165; 171-288)，鹵烷基鹵化物(US4057685)，糖苷鍵(US4673574，US4808700)，己二胺及6-胺基己酸(US4459286)。A possible linker is adipic acid dihydrazide (ADH). Other linkers include B-propionylamino (WO 00/10599), nitrophenyl-ethylamine (Gever et al. (1979) Med. Microbiol. lmmunol. 165; 171-288), halogenated alkyl halides (US4057685), glycosidic bonds (US4673574, US4808700), hexamethylenediamine and 6-aminocaproic acid (US4459286).

在一實施例中，本發明之血清型38醣結合物與載體蛋白直接結合(無需連接子)。In one embodiment, the serotype 38 glycoconjugate of the present invention is directly conjugated to a carrier protein (without a linker).

一般而言，蛋白載體上之以下類型之化學基團可用於偶合/結合： 1) 胺基(例如經由離胺酸)。在一個實施例中，此基團與醣上之羧基直接連接或用例如EDAC (1-乙基-3(3-二甲基胺基丙基)碳二亞胺碳化)之碳化二亞胺化學方法與連接子上之羧基連接。在另一實施例中，此基團與醣上用CDAP或CNBr活化之羥基直接連接或與連接子上之此類基團連接；與具有醛基之醣或連接子連接；與具有丁二醯亞胺基之醣或連接子連接。 2) 羧基(例如經由天冬胺酸或麩胺酸)。在一個實施例中，此基團直接與醣上之胺基連接或用例如EDAC之碳化二亞胺化學方法與連接子上之胺基連接。 3) 巰基(例如經由半胱胺酸)。在一個實施例中，此基團與溴或氯乙醯化醣連接或用順丁烯二醯亞胺化學方法與連接子連接。在一個實施例中，此基團經雙重氮聯苯胺活化/修飾。 4) 羥基(例如經由酪胺酸)。在一個實施例中，此基團經雙重氮聯苯胺活化/修飾。 5) 咪唑基(例如經由組胺酸)。在一個實施例中，此基團經雙重氮聯苯胺活化/修飾。 6) 胍基(例如經由精胺酸)。 7) 吲哚基(例如經由色胺酸)。 In general, the following types of chemical groups on the protein carrier can be used for coupling/conjugation: 1) Amine (e.g., via lysine). In one embodiment, this group is directly linked to a carboxyl group on a sugar or to a carboxyl group on a linker using carbodiimide chemistry such as EDAC (1-ethyl-3(3-dimethylaminopropyl)carbodiimide carbonization). In another embodiment, this group is directly linked to a hydroxyl group on a sugar activated with CDAP or CNBr or to such a group on a linker; to a sugar or linker with an aldehyde group; to a sugar or linker with a succinimidyl group. 2) Carboxyl (e.g., via aspartic acid or glutamine). In one embodiment, this group is directly linked to an amine group on a carbohydrate or is linked to an amine group on a linker using carbodiimide chemistry such as EDAC. 3) Hydroxyl (e.g., via cysteine). In one embodiment, this group is linked to a bromo- or chloroacetylated carbohydrate or is linked to a linker using cis-butylenediimide chemistry. In one embodiment, this group is activated/modified with bisdiazobenzidine. 4) Hydroxyl (e.g., via tyrosine). In one embodiment, this group is activated/modified with bisdiazobenzidine. 5) Imidazolyl (e.g., via histidine). In one embodiment, this group is activated/modified with bisdiazobenzidine. 6) Guanidino (e.g., via arginine). 7) Indole (e.g. via tryptophan).

在血清型38醣上，通常以下基團可用於偶合：OH、COOH或NH2。醛基可在此項技術中已知的不同處理後產生，諸如：過碘酸鹽、酸水解、過氧化氫等。On serotype 38 carbohydrates, the following groups are usually available for coupling: OH, COOH or NH2. Aldehyde groups can be generated by various treatments known in the art, such as periodate, acid hydrolysis, hydrogen peroxide, etc.

在一實施例中，本發明之血清型38醣結合物係使用CDAP化學方法製備。在該實施例中，血清型38醣經1-氰基-4-二甲基胺基吡啶鎓四氟硼酸鹽(CDAP)活化以形成氰酸酯。因此，經活化之醣可直接或經由間隔子(連接子)基團偶合至載體蛋白上之胺基。舉例而言，間隔子可為產生硫醇化多醣之胱胺或半胱胺，該硫醇化多醣可經由在與經順丁烯二醯亞胺活化之載體蛋白(例如，使用N-[γ-順丁烯二醯亞胺基丁醯氧基]丁二醯亞胺酯(GMBS))或經鹵素乙醯化之載體蛋白(例如，使用碘乙醯胺、N-丁二醯亞胺基溴乙酸酯(SBA；SIB)、N-丁二醯亞胺基(4-碘乙醯基)胺基苯甲酸酯(SIAB)、磺基丁二醯亞胺基(4-碘乙醯基)胺基苯甲酸酯(sulfo-SIAB)、N-丁二醯亞胺基碘乙酸酯(SIA)或丁二醯亞胺基3-[溴乙醯胺基]丙酸酯(SBAP))反應之後獲得之硫醚鍵，而與載體偶合。In one embodiment, the serotype 38 carbohydrate conjugate of the present invention is prepared using CDAP chemistry. In this embodiment, serotype 38 carbohydrate is activated by 1-cyano-4-dimethylaminopyridinium tetrafluoroborate (CDAP) to form a cyanate. Thus, the activated carbohydrate can be coupled to an amine group on a carrier protein directly or via a spacer (linker) group. For example, the spacer can be cystamine or cysteamine to produce a thiolated polysaccharide that can be activated by reacting with a carrier protein activated with cis-butylenediamide (e.g., using N-[γ-cis-butylenediamide butyryloxy] succinimide ester (GMBS)) or a carrier protein acetylated with halogen (e.g., using iodoacetamide, N-succinimidyl bromoacetate (SBA; SBA)). The thioether bond obtained after the reaction of succinimidyl (4-iodoacetyl) aminobenzoate (SIAB), sulfosuccinimidyl (4-iodoacetyl) aminobenzoate (sulfo-SIAB), N-succinimidyl iodoacetate (SIA) or succinimidyl 3-[bromoacetylamino] propionate (SBAP)) is coupled with the carrier.

在一較佳實施例中，將經活化之醣的氰酸酯與己二胺或己二酸二醯肼(ADH)偶合，及使用碳化二亞胺(例如EDAC或EDC)化學方法，經由蛋白載體上之羧基，將經胺基衍生之醣與載體蛋白結合。此類結合物描述於例如WO 93/15760、WO 95/08348及WO 96/129094中。In a preferred embodiment, the cyanate of the activated sugar is coupled with hexamethylenediamine or adipic acid dihydrazide (ADH), and the amine-derivatized sugar is conjugated to the carrier protein via the carboxyl groups on the protein carrier using carbodiimide (e.g., EDAC or EDC) chemistry. Such conjugates are described, for example, in WO 93/15760, WO 95/08348, and WO 96/129094.

在一實施例中，使用碳化二亞胺、醯肼、活性酯、降冰片烷、硝基苯甲酸、N-羥基丁二醯亞胺、S-NHS、EDC、TSTU製備本發明之血清型38醣結合物。許多描述於國際專利申請公開案第WO 98/42721號中。結合可涉及可藉由將醣之游離羥基與CDI反應形成的羰基連接子(參見Bethell等人. (1979) 1. Biol. Chern. 254:2572-2574；Hearn等人. (1981) J. Chromatogr. 218:509-518)，接著與蛋白質反應形成胺基甲酸酯鍵。此可涉及將變旋異構端還原成一級羥基，視情況選用之保護/脫除保護該一級羥基，該一級羥基與CDI反應以形成CDI胺基甲酸酯中間物，及將CDI胺基甲酸酯中間物與蛋白質上之胺基偶合。In one embodiment, carbodiimide, hydrazide, active ester, norbornane, nitrobenzoic acid, N-hydroxysuccinimide, S-NHS, EDC, TSTU are used to prepare the serotype 38 glycoconjugates of the present invention. Many are described in International Patent Application Publication No. WO 98/42721. Conjugation may involve a carbonyl linker that can be formed by reacting the free hydroxyl group of the carbohydrate with CDI (see Bethell et al. (1979) I. Biol. Chern. 254:2572-2574; Hearn et al. (1981) J. Chromatogr. 218:509-518), followed by reaction with the protein to form a carbamate bond. This may involve reduction of the mutameric end to a primary hydroxyl group, optionally protecting/deprotecting the primary hydroxyl group, reacting the primary hydroxyl group with CDI to form a CDI carbamate intermediate, and coupling the CDI carbamate intermediate to an amine group on the protein.

直接還原胺化在一實施例中，本發明之血清型38醣結合物藉由直接還原胺化製備(參見例如US 4365170、US 4673574、WO2006/110381、WO2008/079653、WO2008/143709、WO2008/079732、WO2011/110531、WO2012/119972、WO2015110941、WO2015110940、WO2018/144439、WO2018/156491)。 Direct reductive amination In one embodiment, the serotype 38 saccharide conjugate of the present invention is prepared by direct reductive amination (see, e.g., US 4365170, US 4673574, WO2006/110381, WO2008/079653, WO2008/143709, WO2008/079732, WO2011/110531, WO2012/119972, WO2015110941, WO2015110940, WO2018/144439, WO2018/156491).

根據本發明，還原胺化涉及兩個步驟：(1)使血清型38純化之醣氧化(活化)，(2)使經活化之醣及載體蛋白(例如CRM 197、TT或SCP)還原以形成醣結合物。 According to the present invention, reductive amination involves two steps: (1) oxidation (activation) of serotype 38 purified sugars, and (2) reduction of the activated sugars and a carrier protein (eg, CRM 197 , TT or SCP) to form a glycoconjugate.

如上文所提及，在氧化之前，可將血清型38醣之尺寸設定至目標分子量(MW)範圍。因此，在一實施例中，經分離之多醣在氧化之前經尺寸設定。As mentioned above, prior to oxidation, the serotype 38 saccharides can be sized to a target molecular weight (MW) range. Thus, in one embodiment, the isolated polysaccharides are sized prior to oxidation.

在一實施例中，本發明之血清型38醣藉由包含以下步驟之方法與載體蛋白結合： (a)使該血清型38醣與氧化劑反應； (b)使步驟(a)之經活化醣與載體蛋白混配；以及 (c)使該混配之經活化醣及載體蛋白與還原劑反應以形成醣結合物。 In one embodiment, the serotype 38 sugar of the present invention is conjugated to a carrier protein by a method comprising the following steps: (a) reacting the serotype 38 sugar with an oxidizing agent; (b) mixing the activated sugar of step (a) with a carrier protein; and (c) reacting the mixed activated sugar and carrier protein with a reducing agent to form a sugar conjugate.

在一實施例中，本發明之血清型38醣藉由包含以下步驟之方法與載體蛋白結合： (a)使該血清型38醣與氧化劑反應； (a')藉由添加淬滅劑來淬滅該氧化反應； (b)使步驟(a')之經活化醣與載體蛋白混配；以及 (c)使該混配之經活化醣及載體蛋白與還原劑反應以形成醣結合物。 In one embodiment, the serotype 38 sugar of the present invention is conjugated to a carrier protein by a method comprising the following steps: (a) reacting the serotype 38 sugar with an oxidizing agent; (a') quenching the oxidation reaction by adding a quenching agent; (b) mixing the activated sugar of step (a') with a carrier protein; and (c) reacting the mixed activated sugar and carrier protein with a reducing agent to form a sugar conjugate.

在氧化步驟(a)之後，醣稱為經活化的且稱為「經活化醣」。After the oxidation step (a), the sugars are said to be activated and are referred to as "activated sugars."

在一實施例中，氧化劑為將末端羥基氧化成醛之任何氧化劑。在一實施例中，氧化劑為過碘酸鹽。出於本發明之目的，術語「過碘酸鹽」包括過碘酸鹽及過碘酸；術語亦包括偏過碘酸鹽(IO 4 -)及正過碘酸鹽(IO 6 5-)及包括各種過碘酸(例如過碘酸鈉及過碘酸鉀)之鹽。 In one embodiment, the oxidizing agent is any oxidizing agent that oxidizes a terminal hydroxyl group to an aldehyde. In one embodiment, the oxidizing agent is a periodate. For the purposes of the present invention, the term "periodate" includes periodates and periodic acid; the term also includes metaperiodates (IO 4 - ) and orthoperiodates (IO 6 5- ) and salts including various periodic acids (e.g., sodium periodate and potassium periodate).

在一實施例中，氧化劑在二價陽離子存在下為過碘酸鹽(參見WO2008/143709)。In one embodiment, the oxidizing agent is a periodate in the presence of a divalent cation (see WO2008/143709).

在一實施例中，氧化劑為過碘酸。在一實施例中，氧化劑在二價陽離子存在下為過碘酸。在一實施例中，氧化劑在Mg 2+存在下為過碘酸。在一實施例中，氧化劑在Ca 2+存在下為過碘酸。在一實施例中，氧化劑為正過碘酸鹽。 In one embodiment, the oxidant is periodic acid. In one embodiment, the oxidant is periodic acid in the presence of divalent cations. In one embodiment, the oxidant is periodic acid in the presence of Mg 2+ . In one embodiment, the oxidant is periodic acid in the presence of Ca 2+ . In one embodiment, the oxidant is orthoperiodate.

在一個較佳實施例中，氧化劑為過碘酸鈉。在一實施例中，用於氧化之過碘酸鹽係偏過碘酸鹽。在一實施例中，用於氧化之過碘酸鹽係偏過碘酸鈉。In a preferred embodiment, the oxidizing agent is sodium periodate. In one embodiment, the periodate salt used for oxidation is metaperiodate. In one embodiment, the periodate salt used for oxidation is sodium metaperiodate.

當多醣與過碘酸鹽反應時，過碘酸鹽使鄰位羥基氧化以形成羰基或醛基，且引起C-C鍵裂解。出於此原因，「使多醣與過碘酸鹽反應」之術語包括藉由過碘酸鹽氧化鄰位羥基。When a polysaccharide is reacted with a periodate salt, the periodate salt oxidizes the vicinal hydroxyl group to form a carbonyl group or an aldehyde group and causes cleavage of the C-C bond. For this reason, the term "reacting a polysaccharide with a periodate salt" includes oxidation of the vicinal hydroxyl group by the periodate salt.

在一個實施例中，步驟a)包含使多醣與0.01至2莫耳當量之過碘酸鹽反應。在一個實施例中，步驟a)包含使多醣與0.1至1.0莫耳當量之過碘酸鹽反應。在一個實施例中，步驟a)包含使多醣與0.1至0.5莫耳當量之過碘酸鹽反應。In one embodiment, step a) comprises reacting the polysaccharide with 0.01 to 2 molar equivalents of a periodate salt. In one embodiment, step a) comprises reacting the polysaccharide with 0.1 to 1.0 molar equivalents of a periodate salt. In one embodiment, step a) comprises reacting the polysaccharide with 0.1 to 0.5 molar equivalents of a periodate salt.

在一實施例中，氧化劑為穩定硝醯基自由基化合物與氧化劑之混合物(參見WO2014097099)。In one embodiment, the oxidizing agent is a mixture of a stable nitroxyl radical compound and an oxidizing agent (see WO2014097099).

在一態樣中，該穩定硝醯基自由基化合物為帶有TEMPO或PROXYL (2,2,5,5-四甲基-1-吡咯啶基氧基)部分之分子。較佳地，該分子能夠在氧化劑存在下選擇性地氧化一級醇，以產生醛基，而不影響二級羥基。更佳地，該分子能夠在氧化劑存在下選擇性地氧化一級醇，以產生醛基，而不過度氧化成羧基。在一態樣中，該穩定硝醯基自由基化合物為TEMPO、2,2,6,6-四甲基-4-(甲基磺醯氧基)-1-哌啶氧基、4-膦醯氧基-TEMPO、4-側氧基-TEMPO、4-甲氧基-TEMPO、4-異硫氰基-TEMPO、4-(2-碘乙醯胺基)-TEMPO自由基、4-羥基-TEMPO、4-氰基-TEMPO、4-羧基-TEMPO、4-(2-溴乙醯胺基)-TEMPO或4-胺基-TEMPO、4-乙醯胺基-2,2,6,6-四甲基哌啶1-氧基。較佳地，該穩定硝醯基自由基化合物為TEMPO。在一態樣中，該穩定硝醯基自由基化合物係選自由以下組成之群：TEMPO、2,2,6,6-四甲基-4-(甲基磺醯氧基)-1-哌啶氧基、4-膦醯氧基-TEMPO、4-側氧基-TEMPO、4-甲氧基-TEMPO、4-異硫氰基-TEMPO、4-(2-碘乙醯胺基)-TEMPO自由基、4-羥基-TEMPO、4-氰基-TEMPO、4-羧基-TEMPO、4-(2-溴乙醯胺基)-TEMPO、4-胺基-TEMPO、4-乙醯胺基-2,2,6,6-四甲基哌啶1-氧基。較佳地，該穩定硝醯基自由基化合物為TEMPO。在另一態樣中，該穩定硝醯基自由基化合物為3β-DOXYL-5α-膽甾烷、5-DOXYL-硬脂酸、16-DOXYL-硬脂酸、5-DOXYL-硬脂酸甲酯、3-(胺基甲基)-PROXYL、3-胺甲醯基-PROXYL、3-胺甲醯基-2,2,5,5-四甲基-3-吡咯啉-1-氧基、3-羧基-PROXYL或3-氰基-PROXYL。在另一態樣中，該穩定硝醯基自由基化合物係選自由以下組成之群：3β-DOXYL-5α-膽甾烷、5-DOXYL-硬脂酸、16-DOXYL-硬脂酸、5-DOXYL-硬脂酸甲酯、3-(胺基甲基)-PROXYL、3-胺甲醯基-PROXYL、3-胺甲醯基-2,2,5,5-四甲基-3-吡咯啉-1-氧基、3-羧基-PROXYL、3-氰基-PROXYL。In one embodiment, the stable nitro radical compound is a molecule with a TEMPO or PROXYL (2,2,5,5-tetramethyl-1-pyrrolidinyloxy) moiety. Preferably, the molecule is capable of selectively oxidizing a primary alcohol in the presence of an oxidizing agent to generate an aldehyde group without affecting a secondary hydroxyl group. More preferably, the molecule is capable of selectively oxidizing a primary alcohol in the presence of an oxidizing agent to generate an aldehyde group without excessive oxidation to a carboxyl group. In one embodiment, the stable nitro radical compound is TEMPO, 2,2,6,6-tetramethyl-4-(methylsulfonyloxy)-1-piperidinyloxy, 4-phosphonyloxy-TEMPO, 4-oxo-TEMPO, 4-methoxy-TEMPO, 4-isothiocyanato-TEMPO, 4-(2-iodoacetamido)-TEMPO radical, 4-hydroxy-TEMPO, 4-cyano-TEMPO, 4-carboxyl-TEMPO, 4-(2-bromoacetamido)-TEMPO or 4-amino-TEMPO, 4-acetamido-2,2,6,6-tetramethylpiperidinyl-1-oxyl. Preferably, the stable nitro radical compound is TEMPO. In one embodiment, the stable nitro radical compound is selected from the group consisting of TEMPO, 2,2,6,6-tetramethyl-4-(methylsulfonyloxy)-1-piperidinyloxy, 4-phosphonyloxy-TEMPO, 4-oxo-TEMPO, 4-methoxy-TEMPO, 4-isothiocyanato-TEMPO, 4-(2-iodoacetamido)-TEMPO radical, 4-hydroxy-TEMPO, 4-cyano-TEMPO, 4-carboxyl-TEMPO, 4-(2-bromoacetamido)-TEMPO, 4-amino-TEMPO, 4-acetamido-2,2,6,6-tetramethylpiperidinyl-1-oxyl. Preferably, the stable nitro radical compound is TEMPO. In another embodiment, the stable nitro radical compound is 3β-DOXYL-5α-cholestane, 5-DOXYL-stearic acid, 16-DOXYL-stearic acid, 5-DOXYL-stearate methyl ester, 3-(aminomethyl)-PROXYL, 3-aminoformyl-PROXYL, 3-aminoformyl-2,2,5,5-tetramethyl-3-pyrroline-1-oxyl, 3-carboxy-PROXYL or 3-cyano-PROXYL. In another aspect, the stable nitro radical compound is selected from the group consisting of 3β-DOXYL-5α-cholestane, 5-DOXYL-stearic acid, 16-DOXYL-stearic acid, 5-DOXYL-stearate methyl ester, 3-(aminomethyl)-PROXYL, 3-aminoformyl-PROXYL, 3-aminoformyl-2,2,5,5-tetramethyl-3-pyrroline-1-oxyl, 3-carboxyl-PROXYL, 3-cyano-PROXYL.

在一態樣中，氧化劑為帶有N-鹵基部分之分子。較佳地，該分子能夠在硝醯基自由基化合物存在下選擇性氧化一級醇。在一態樣中，該氧化劑為N-氯代丁二醯亞胺、N-溴代丁二醯亞胺、N-碘丁二醯亞胺、二氯異三聚氰酸、1,3,5-三氯-1,3,5-三𠯤烷-2,4,6-三酮、二溴異三聚氰酸、1,3,5-三溴-1,3,5-三𠯤烷-2,4,6-三酮、二碘異三聚氰酸或1,3,5-三碘-1,3,5-三𠯤烷-2,4,6-三酮。在一態樣中，該氧化劑係選自由以下組成之群：N-氯代丁二醯亞胺、N-溴代丁二醯亞胺、N-碘丁二醯亞胺、二氯異三聚氰酸、1,3,5-三氯-1,3,5-三𠯤烷-2,4,6-三酮、二溴異三聚氰酸、1,3,5-三溴-1,3,5-三𠯤烷-2,4,6-三酮、二碘異三聚氰酸及1,3,5-三碘-1,3,5-三𠯤烷-2,4,6-三酮。較佳地，該氧化劑為N-氯代丁二醯亞胺。In one embodiment, the oxidant is a molecule with an N-halogen moiety. Preferably, the molecule is capable of selectively oxidizing a primary alcohol in the presence of a nitro radical compound. In one embodiment, the oxidant is N-chlorosuccinimide, N-bromosuccinimide, N-iodosuccinimide, dichloroisocyanuric acid, 1,3,5-trichloro-1,3,5-trioxane-2,4,6-trione, dibromoisocyanuric acid, 1,3,5-tribromo-1,3,5-trioxane-2,4,6-trione, diiodoisocyanuric acid or 1,3,5-triiodo-1,3,5-trioxane-2,4,6-trione. In one embodiment, the oxidant is selected from the group consisting of: N-chlorosuccinimide, N-bromosuccinimide, N-iodosuccinimide, dichloroisocyanuric acid, 1,3,5-trichloro-1,3,5-trioxane-2,4,6-trione, dibromoisocyanuric acid, 1,3,5-tribromo-1,3,5-trioxane-2,4,6-trione, diiodoisocyanuric acid and 1,3,5-triiodo-1,3,5-trioxane-2,4,6-trione. Preferably, the oxidant is N-chlorosuccinimide.

在一態樣中，該穩定硝醯基自由基化合物為2,2,6,6-四甲基-1-哌啶基氧基自由基(TEMPO)，且該氧化劑為N-氯代丁二醯亞胺(NCS)。In one embodiment, the stable nitroxyl radical compound is 2,2,6,6-tetramethyl-1-piperidinyloxy radical (TEMPO), and the oxidizing agent is N-chlorosuccinimide (NCS).

在一個實施例中，步驟a')之淬滅劑係選自鄰二醇、1,2-胺基醇、胺基酸、麩胱甘肽、亞硫酸鹽、硫酸氫鹽、二亞硫酸鹽、偏亞硫酸氫鹽、硫代硫酸鹽、亞磷酸鹽、次磷酸鹽或亞磷酸。In one embodiment, the quenching agent of step a') is selected from a vicinal diol, a 1,2-amino alcohol, an amino acid, glutathione, a sulfite, a bisulfate, a disulfite, a metabisulfite, a thiosulfate, a phosphite, a hypophosphite or a phosphorous acid.

在一個實施例中，淬滅劑為式(I)之1,2-胺基醇： 其中R 1係選自H、甲基、乙基、丙基或異丙基。 In one embodiment, the quencher is a 1,2-amino alcohol of formula (I): wherein R 1 is selected from H, methyl, ethyl, propyl or isopropyl.

在一個實施例中，淬滅劑係選自亞硫酸鹽、硫酸氫鹽、二亞硫酸鹽、偏亞硫酸氫鹽、硫代硫酸鹽、亞磷酸鹽、次磷酸鹽或亞磷酸之鈉鹽及鉀鹽。In one embodiment, the quencher is selected from the sodium and potassium salts of sulfites, bisulfites, disulfites, metabisulfites, thiosulfates, phosphites, hypophosphites, or phosphites.

在一個實施例中，淬滅劑為胺基酸。在此類實施例中，該胺基酸可選自絲胺酸、蘇胺酸、半胱胺酸、胱胺酸、甲硫胺酸、脯胺酸、羥脯胺酸、色胺酸、酪胺酸及組胺酸。In one embodiment, the quencher is an amino acid. In such embodiments, the amino acid can be selected from serine, threonine, cysteine, cystine, methionine, proline, hydroxyproline, tryptophan, tyrosine and histidine.

在一個實施例中，淬滅劑係亞硫酸鹽，諸如硫酸氫鹽、二亞硫磺酸、偏亞硫酸氫鹽、硫代硫酸鹽。In one embodiment, the quenching agent is a sulfite, such as bisulfate, disulfurous acid, metabisulfite, thiosulfate.

在一個實施例中，淬滅劑為包含兩個鄰位羥基(鄰二醇)之化合物，亦即共價連接至兩個相鄰碳原子之兩個羥基。In one embodiment, the quencher is a compound comprising two vicinal hydroxyl groups (vicinal diol), i.e., two hydroxyl groups covalently linked to two adjacent carbon atoms.

較佳地，淬滅劑為式(II)之化合物： 其中R 1及R 2各自獨立地係選自H、甲基、乙基、丙基或異丙基。 Preferably, the quencher is a compound of formula (II): wherein R1 and R2 are each independently selected from H, methyl, ethyl, propyl or isopropyl.

在較佳實施例中，淬滅劑為丙三醇、乙二醇、丙-1,2-二醇、丁-1,2-二醇或丁-2,3-二醇或抗壞血酸。在一甚至較佳實施例中，淬滅劑係丁-2,3-二醇。In a preferred embodiment, the quencher is glycerol, ethylene glycol, propane-1,2-diol, butane-1,2-diol or butane-2,3-diol or ascorbic acid. In an even more preferred embodiment, the quencher is butane-2,3-diol.

在一較佳實施例中，經活化血清型38醣的氧化程度(在本文件中亦被稱為「活化程度」)在2與30之間。在一實施例中，經活化血清型38多醣之氧化程度(DO)在10與25之間。In a preferred embodiment, the degree of oxidation (also referred to herein as "degree of activation") of the activated serotype 38 saccharide is between 2 and 30. In one embodiment, the degree of oxidation (DO) of the activated serotype 38 polysaccharide is between 10 and 25.

在一個實施例中，經活化之醣及載體蛋白在步驟b)之前凍乾。In one embodiment, the activated sugars and carrier protein are lyophilized before step b).

在一實施例中，步驟b)處之經活化之血清型38醣與載體蛋白之初始輸入比率(重量/重量)在4:1與0.1:1之間。在一實施例中，步驟b)處之經活化之血清型38醣與載體蛋白之初始輸入比率(重量/重量)在1.5:1與0.5:1之間。In one embodiment, the initial input ratio (weight/weight) of activated serotype 38 saccharide to carrier protein at step b) is between 4: 1 and 0.1: 1. In one embodiment, the initial input ratio (weight/weight) of activated serotype 38 saccharide to carrier protein at step b) is between 1.5: 1 and 0.5: 1.

在一實施例中，還原反應(c)在水溶劑中進行。在另一實施例中，還原反應(c)在非質子性溶劑中進行。In one embodiment, the reduction reaction (c) is carried out in an aqueous solvent. In another embodiment, the reduction reaction (c) is carried out in an aprotic solvent.

在一實施例中，還原反應(c)在二甲亞碸(DMSO)或二甲基甲醯胺(DMF)存在下進行。在一實施例中，還原反應(c)在二甲基甲醯胺(DMF)存在下進行。在一實施例中，還原反應(c)在二甲亞碸(DMSO)存在下進行。In one embodiment, the reduction reaction (c) is carried out in the presence of dimethyl sulfoxide (DMSO) or dimethylformamide (DMF). In one embodiment, the reduction reaction (c) is carried out in the presence of dimethylformamide (DMF). In one embodiment, the reduction reaction (c) is carried out in the presence of dimethyl sulfoxide (DMSO).

在一個實施例中，還原反應(c)在基本上由二甲亞碸(DMSO)或二甲基甲醯胺(DMF)組成之溶液中進行。在一個實施例中，還原反應(c)在基本上由二甲基甲醯胺(DMF)組成之溶液中進行。在一個實施例中，還原反應(c)在基本上由二甲亞碸(DMSO)組成之溶液中進行。In one embodiment, the reduction reaction (c) is carried out in a solution consisting essentially of dimethyl sulfoxide (DMSO) or dimethylformamide (DMF). In one embodiment, the reduction reaction (c) is carried out in a solution consisting essentially of dimethylformamide (DMF). In one embodiment, the reduction reaction (c) is carried out in a solution consisting essentially of dimethyl sulfoxide (DMSO).

在一實施例中，還原反應(c)在DMSO (二甲亞碸)或DMF (二甲基甲醯胺)溶劑中進行。在一實施例中，還原反應(c)在DMSO (二甲亞碸)溶劑中進行。In one embodiment, the reduction reaction (c) is carried out in DMSO (dimethyl sulfoxide) or DMF (dimethylformamide) solvent. In one embodiment, the reduction reaction (c) is carried out in DMSO (dimethyl sulfoxide) solvent.

在一實施例中，還原劑為氰基硼氫化鈉、三乙醯氧基硼氫化鈉、在布朗斯特酸(Bronsted acid)或路易斯酸(Lewis acid)存在下之硼氫化鈉或硼氫化鋅、胺硼烷，諸如吡啶硼烷、2-甲吡啶硼烷、2,6-二硼烷-甲醇、二甲胺-硼烷、t-BuMe iPrN-BH 3、苯甲胺-BH 3或5-乙基-2-甲基吡啶硼烷(PEMB)。在一實施例中，還原劑為三乙醯氧基硼氫化鈉。在一較佳實施例中，還原劑為氰基硼氫化鈉。在一實施例中，還原劑在鎳存在下為氰基硼氫化鈉(參見WO2018144439)。 In one embodiment, the reducing agent is sodium cyanoborohydride, sodium triacetoxyborohydride, sodium borohydride or zinc borohydride in the presence of Bronsted acid or Lewis acid, amine borane, such as pyridine borane, 2-methylpyridine borane, 2,6-diborane-methanol, dimethylamine-borane, t-BuMe i PrN-BH 3 , benzylamine-BH 3 or 5-ethyl-2-methylpyridine borane (PEMB). In one embodiment, the reducing agent is sodium triacetoxyborohydride. In a preferred embodiment, the reducing agent is sodium cyanoborohydride. In one embodiment, the reducing agent is sodium cyanoborohydride in the presence of nickel (see WO2018144439).

在一個實施例中，在步驟c)處使用0.2與20莫耳當量之間的還原劑。在一個實施例中，在步驟c)處使用0.5與10莫耳當量之間的還原劑。在一個實施例中，在步驟c)處使用1.0與5莫耳當量之間的還原劑。In one embodiment, between 0.2 and 20 molar equivalents of reducing agent are used at step c). In one embodiment, between 0.5 and 10 molar equivalents of reducing agent are used at step c). In one embodiment, between 1.0 and 5 molar equivalents of reducing agent are used at step c).

在還原反應結束時，結合物中可存在剩餘未反應之醛基，可使用適合的封端劑對此等醛基進行封端。在一個實施例中，此封端劑為硼氫化鈉(NaBH 4)。在一實施例中，封端藉由將步驟c)之產物與1至20莫耳當量之硼氫化鈉混合來達成。在一實施例中，封端藉由將步驟c)之產物與1至10莫耳當量之硼氫化鈉混合來達成。在一實施例中，封端藉由將步驟c)之產物與1至5莫耳當量之硼氫化鈉混合來達成。 At the end of the reduction reaction, there may be residual unreacted aldehyde groups in the conjugate, and these aldehyde groups can be capped using a suitable capping agent. In one embodiment, the capping agent is sodium borohydride (NaBH 4 ). In one embodiment, the capping is achieved by mixing the product of step c) with 1 to 20 molar equivalents of sodium borohydride. In one embodiment, the capping is achieved by mixing the product of step c) with 1 to 10 molar equivalents of sodium borohydride. In one embodiment, the capping is achieved by mixing the product of step c) with 1 to 5 molar equivalents of sodium borohydride.

CDI 及 / 或 CDT 化學方法在一實施例中，本發明之血清型38醣結合物係藉由WO2022249107中所揭露之CDI及/或CDT化學方法來製備。 CDI and / or CDT chemical methods In one embodiment, the serotype 38 saccharide conjugate of the present invention is prepared by the CDI and/or CDT chemical methods disclosed in WO2022249107.

CDI及/或CDT化學方法涉及兩個步驟：(1)使血清型38醣與CDI及/或CDT在非質子性溶劑中反應以產生經活化醣(活化)，(2)使經活化醣與載體蛋白質(例如CRM 197、TT或SCP)反應以形成醣結合物。 The CDI and/or CDT chemistry involves two steps: (1) reacting serotype 38 saccharides with CDI and/or CDT in an aprotic solvent to produce activated saccharides (activation), and (2) reacting the activated saccharides with a carrier protein (eg, CRM 197 , TT or SCP) to form a glycoconjugate.

在一實施例中，步驟(1)之活化劑為1,1'-羰基二咪唑(CDI)。在一實施例中，步驟(1)之活化劑為1,1'-羰基-二-(1,2,4-三唑) (CDT)。In one embodiment, the activating agent in step (1) is 1,1'-carbonyldiimidazole (CDI). In one embodiment, the activating agent in step (1) is 1,1'-carbonyl-di-(1,2,4-triazole) (CDT).

如上文所提及，在用CDI及/或CDT活化之前，可將血清型38醣之尺寸設定至目標分子量(MW)範圍。As mentioned above, serotype 38 saccharides can be sized to a target molecular weight (MW) range prior to activation with CDI and/or CDT.

因此，在一實施例中，在用CDI活化之前，血清型38醣經尺寸設定。在一實施例中，在用CDT活化之前，經分離之多醣經尺寸設定。在一實施例中，血清型38醣經尺寸設定為上文所定義之目標分子量(MW)範圍中之任一者。Thus, in one embodiment, serotype 38 carbohydrates are sized prior to activation with CDI. In one embodiment, the isolated polysaccharides are sized prior to activation with CDT. In one embodiment, serotype 38 carbohydrates are sized to any of the target molecular weight (MW) ranges defined above.

因此，在一實施例中，血清型38醣藉由包含以下步驟之方法與載體蛋白結合： (a)使該經分離多醣與CDI及/或CDT在非質子性溶劑中反應； (b)使步驟(a)之該經活化多醣與載體蛋白在非質子性溶劑中反應以形成醣結合物。 Therefore, in one embodiment, serotype 38 saccharide is conjugated to a carrier protein by a method comprising the following steps: (a) reacting the separated polysaccharide with CDI and/or CDT in an aprotic solvent; (b) reacting the activated polysaccharide of step (a) with a carrier protein in an aprotic solvent to form a saccharide conjugate.

在步驟(a)之後，多醣被稱為經活化的且稱為「經活化之多醣」。After step (a), the polysaccharide is said to be activated and is referred to as "activated polysaccharide".

在一個實施例中，步驟a)包含使血清型38醣與CDI反應。In one embodiment, step a) comprises reacting serotype 38 saccharide with CDI.

在一個實施例中，步驟a)包含使血清型38醣與相對於反應混合物中所存在之血清型38醣的量在0.5至10莫耳當量之間的量的CDI反應。In one embodiment, step a) comprises reacting serotype 38 saccharide with CDI in an amount between 0.5 and 10 molar equivalents relative to the amount of serotype 38 saccharide present in the reaction mixture.

在一個實施例中，步驟a)包含使血清型38醣與CDT反應。In one embodiment, step a) comprises reacting serotype 38 saccharide with CDT.

在一個實施例中，步驟a)包含使血清型38醣與相對於反應混合物中所存在之血清型38醣的量在0.5至10莫耳當量之間的量的CDT反應。In one embodiment, step a) comprises reacting serotype 38 saccharide with CDT in an amount between 0.5 and 10 molar equivalents relative to the amount of serotype 38 saccharide present in the reaction mixture.

在一實施例中，活化反應a)在二甲亞碸(DMSO)、二甲基甲醯胺(DMF)、二甲基乙醯胺、N-甲基-2-吡咯啶酮或六甲基磷醯胺(HMPA)存在下進行。在一個實施例中，活化反應a)在二甲亞碸(DMSO)存在下進行。In one embodiment, the activation reaction a) is carried out in the presence of dimethyl sulfoxide (DMSO), dimethylformamide (DMF), dimethylacetamide, N-methyl-2-pyrrolidone or hexamethylphosphatamide (HMPA). In one embodiment, the activation reaction a) is carried out in the presence of dimethyl sulfoxide (DMSO).

在一個實施例中，活化反應a)在基本上由二甲亞碸(DMSO)或二甲基甲醯胺(DMF)組成之溶液中進行。在一個實施例中，活化反應a)在基本上由二甲亞碸(DMSO)組成之溶液中進行。In one embodiment, the activation reaction a) is carried out in a solution consisting essentially of dimethyl sulfoxide (DMSO) or dimethylformamide (DMF). In one embodiment, the activation reaction a) is carried out in a solution consisting essentially of dimethyl sulfoxide (DMSO).

在一實施例中，結合反應b)在二甲亞碸(DMSO)、二甲基甲醯胺(DMF)、二甲基乙醯胺、N-甲基-2-吡咯啶酮或六甲基磷醯胺(HMPA)存在下進行。In one embodiment, the conjugation reaction b) is carried out in the presence of dimethyl sulfoxide (DMSO), dimethylformamide (DMF), dimethylacetamide, N-methyl-2-pyrrolidone or hexamethylphosphatide (HMPA).

在一實施例中，結合反應b)在二甲亞碸(DMSO)存在下進行。In one embodiment, the conjugation reaction b) is carried out in the presence of dimethyl sulfoxide (DMSO).

在一個實施例中，結合反應b)在基本上由二甲亞碸(DMSO)或二甲基甲醯胺(DMF)組成之溶液中進行。在一個實施例中，結合反應b)在基本上由二甲亞碸(DMSO)組成之溶液中進行。In one embodiment, the conjugation reaction b) is carried out in a solution consisting essentially of dimethyl sulfoxide (DMSO) or dimethylformamide (DMF). In one embodiment, the conjugation reaction b) is carried out in a solution consisting essentially of dimethyl sulfoxide (DMSO).

在一個實施例中，弱有機鹼可在活化反應a)之後但在結合反應b)之前添加至反應混合物中。可在載體蛋白引入反應混合物之前或之後添加弱有機鹼。因此，在一個實施例中，在引入載體蛋白之前，將弱有機鹼添加至反應混合物中。在另一實施例中，在引入載體蛋白之後，將弱有機鹼添加至反應混合物中。弱有機鹼可選自烷胺、咪唑、三唑、吡啶、組胺酸及胍。烷胺包括烷基一級胺，諸如甲胺、乙胺、丙胺、異丙胺；烷基二級胺，諸如二甲胺、二乙胺、二丙胺、二異丙胺；烷基三級胺，諸如三甲胺、三乙胺、三異丙胺、二-N,N'-異丙基乙胺等。在一實施例中，弱有機鹼為烷胺。在一實施例中，弱有機鹼為咪唑。在一實施例中，弱有機鹼為三唑。在一實施例中，弱有機鹼為吡啶。在一實施例中，弱有機鹼為組胺酸。在一實施例中，弱有機鹼為胍。In one embodiment, the weak organic base can be added to the reaction mixture after the activation reaction a) but before the binding reaction b). The weak organic base can be added before or after the carrier protein is introduced into the reaction mixture. Therefore, in one embodiment, the weak organic base is added to the reaction mixture before the carrier protein is introduced. In another embodiment, the weak organic base is added to the reaction mixture after the carrier protein is introduced. The weak organic base can be selected from alkylamines, imidazoles, triazoles, pyridines, histidine and guanidines. Alkylamines include alkyl primary amines, such as methylamine, ethylamine, propylamine, isopropylamine; alkyl diamines, such as dimethylamine, diethylamine, dipropylamine, diisopropylamine; alkyl tertiary amines, such as trimethylamine, triethylamine, triisopropylamine, di-N,N'-isopropylethylamine, etc. In one embodiment, the weak organic base is an alkylamine. In one embodiment, the weak organic base is imidazole. In one embodiment, the weak organic base is triazole. In one embodiment, the weak organic base is pyridine. In one embodiment, the weak organic base is histidine. In one embodiment, the weak organic base is guanidine.

在一個實施例中，在結合反應b)後，經活化多醣之未結合的反應性位點被水解。在一個實施例中，藉由向結合溶液中添加水溶液來水解未結合之反應性位點。在一個實施例中，藉由向結合溶液中添加水性緩衝溶液來水解未結合之反應性位點。在一個實施例中，藉由向結合溶液中添加水性緩衝溶液且將pH值調節至約3.0至約10.0之間來水解未結合之反應性位點。在一個實施例中，藉由向結合溶液中添加水性緩衝溶液且將pH值調節至約7.0至約10.0之間來水解未結合之反應性位點。在一個實施例中，藉由向結合溶液中添加水性緩衝溶液且將pH值調節至約3.0至約7.0之間來水解未結合之反應性位點。在一個實施例中，藉由向結合溶液中添加水性緩衝溶液且將pH值調節至約4.0來水解未結合之反應性位點。在一個實施例中，藉由向結合溶液中添加水性緩衝溶液且將pH值調節至約9.0來水解未結合之反應性位點。In one embodiment, after the binding reaction b), the unbound reactive sites of the activated polysaccharide are hydrolyzed. In one embodiment, the unbound reactive sites are hydrolyzed by adding an aqueous solution to the binding solution. In one embodiment, the unbound reactive sites are hydrolyzed by adding an aqueous buffer solution to the binding solution. In one embodiment, the unbound reactive sites are hydrolyzed by adding an aqueous buffer solution to the binding solution and adjusting the pH to between about 3.0 and about 10.0. In one embodiment, the unbound reactive sites are hydrolyzed by adding an aqueous buffer solution to the binding solution and adjusting the pH to between about 7.0 and about 10.0. In one embodiment, unbound reactive sites are hydrolyzed by adding an aqueous buffer solution to the binding solution and adjusting the pH to between about 3.0 and about 7.0. In one embodiment, unbound reactive sites are hydrolyzed by adding an aqueous buffer solution to the binding solution and adjusting the pH to about 4.0. In one embodiment, unbound reactive sites are hydrolyzed by adding an aqueous buffer solution to the binding solution and adjusting the pH to about 9.0.

eTEC 化學方法在一實施例中，本發明之血清型38醣結合物係藉由WO2014027302中所揭露之eTEC化學方法來製備。 eTEC Chemical Method In one embodiment, the serotype 38 saccharide conjugate of the present invention is prepared by the eTEC chemical method disclosed in WO2014027302.

eTEC間隔子包括七個線性原子(亦即-C(O)NH(CH2)2SCH2C(O)-)，且在醣與載體蛋白之間提供穩定硫醚及醯胺鍵。eTEC連接之醣結合物的合成涉及醣之經活化羥基與硫烷基胺試劑之胺基(例如，胱胺或半胱胺或其鹽)反應，從而形成與醣之胺基甲酸酯鍵以提供經硫醇化醣。產生一或多個游離硫氫基係藉由與還原劑反應以提供經活化硫醇化醣而實現。經活化硫醇化醣之游離硫氫基與含胺之殘基上具有一或多個α-鹵乙醯胺基之經活化載體蛋白之反應產生硫醚鍵以形成結合物，其中載體蛋白經由醯胺鍵連接至eTEC間隔子。The eTEC spacer comprises seven linear atoms (i.e., -C(O)NH(CH2)2SCH2C(O)-) and provides stable thioether and amide bonds between the sugar and the carrier protein. The synthesis of eTEC-linked glycoconjugates involves the reaction of activated hydroxyl groups of the sugar with the amine group of a sulfanylamine reagent (e.g., cystamine or cysteamine or a salt thereof) to form a carbamate bond with the sugar to provide the thiolated sugar. Generation of one or more free sulfhydryl groups is achieved by reaction with a reducing agent to provide the activated thiolated sugar. The reaction of the free sulfhydryl group of the activated thiolated sugar with the activated carrier protein having one or more α-haloacetamide groups on the amine-containing residue generates a thioether bond to form a conjugate, wherein the carrier protein is linked to the eTEC spacer via the amide bond.

因此，在一個實施例中，本發明之血清型38醣結合物包含透過胺基甲酸(2-((2-側氧基乙基)硫基)乙基)酯(eTEC)間隔子與載體蛋白共價結合之血清型38醣。Therefore, in one embodiment, the serotype 38 saccharide conjugate of the present invention comprises serotype 38 saccharide covalently bound to a carrier protein via an eTEC spacer.

在一實施例中，本發明之血清型38醣結合物包含透過胺基甲酸(2-((2-側氧基乙基)硫基)乙基)酯(eTEC)間隔子與載體蛋白結合之血清型38醣，其中該醣透過胺基甲酸酯鍵與該eTEC間隔子共價連接，且其中該載體蛋白透過醯胺鍵與該eTEC間隔子共價連接。In one embodiment, the serotype 38 carbohydrate conjugate of the invention comprises a serotype 38 carbohydrate conjugated to a carrier protein via a (2-((2-hydroxyethyl)thio)ethyl) carbamate (eTEC) spacer, wherein the carbohydrate is covalently linked to the eTEC spacer via a carbamate bond, and wherein the carrier protein is covalently linked to the eTEC spacer via an amide bond.

本發明之eTEC連接的醣結合物可由以下通式(III)表示： (III)，其中(醣)表示血清型38醣。 The eTEC-linked carbohydrate conjugate of the present invention can be represented by the following general formula (III): (III), wherein (sugar) represents serotype 38 sugar.

式(III)為本發明之醣結合物的示意性表示。不應理解為醣與載體蛋白之間僅存在一個鍵。實際上，個別載體蛋白(CP)分子可連接至多於一個血清型38醣分子，且個體醣分子可連接至多於一個個別載體蛋白(CP)分子。另外，大部分醣重複單元保持未經修飾，且少數醣重複單元處存在載體蛋白與醣之共價連接。Formula (III) is a schematic representation of the glycoconjugate of the present invention. It should not be understood that there is only one bond between the carbohydrate and the carrier protein. In fact, an individual carrier protein (CP) molecule can be linked to more than one serotype 38 carbohydrate molecule, and an individual carbohydrate molecule can be linked to more than one individual carrier protein (CP) molecule. In addition, most of the carbohydrate repeat units remain unmodified, and a covalent bond between the carrier protein and the carbohydrate exists at a few carbohydrate repeat units.

點擊化學在一較佳實施例中，本發明之血清型38醣結合物係藉由點擊化學製備(參見例如PCT/IB2023/050202)。 Click Chemistry In a preferred embodiment, the serotype 38 carbohydrate conjugate of the present invention is prepared by click chemistry (see, e.g., PCT/IB2023/050202).

因此，在一實施例中，本發明之血清型38醣結合物包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(IV)： ， 其中X係選自由CH 2(CH 2) n '、(CH 2CH 2O) mCH 2CH 2、NHCO(CH 2) n '、NHCO(CH 2CH 2O) mCH 2CH 2、OCH 2(CH 2) n '及O(CH 2CH 2O) mCH 2CH 2組成之群；其中n'係選自1至10且m係選自1至4， 其中X'係選自由CH 2O(CH 2) n''CH 2C=O、CH 2O(CH 2CH 2O) m'(CH 2) n''CH 2C=O組成之群，其中n''係選自0至10且m'係選自0至4， 其中方括號中之結構表示該血清型38醣之重複單元，且其中n表示重複單元之數目。 Therefore, in one embodiment, the serotype 38 saccharide conjugate of the present invention comprises a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and has the general formula (IV): , wherein X is selected from the group consisting of CH 2 (CH 2 ) n ′ , (CH 2 CH 2 O) m CH 2 CH 2 , NHCO(CH 2 ) n ′ , NHCO(CH 2 CH 2 O) m CH 2 CH 2 , OCH 2 (CH 2 ) n ′ , and O(CH 2 CH 2 O) m CH 2 CH 2 ; wherein n′ is selected from 1 to 10 and m is selected from 1 to 4, wherein X′ is selected from the group consisting of CH 2 O(CH 2 ) n′ CH 2 C═O, CH 2 O(CH 2 CH 2 O) m′ (CH 2 ) n′ CH 2 C═O, wherein n′ is selected from 0 to 10 and m′ is selected from 0 to 4, wherein the structure in square brackets represents the repeating unit of the serotype 38 carbohydrate, and wherein n represents the number of repeating units.

在一特定態樣中，本發明係關於一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(IV)，其中X為CH 2(CH 2) n '，其中n'為2，且其中X'為CH 2O(CH 2) n ''CH 2C=O，其中n''為1。 In a specific embodiment, the present invention relates to a serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (IV), wherein X is CH2 ( CH2 ) n ' , wherein n' is 2, and wherein X' is CH2O ( CH2 ) n''CH2C =O, wherein n'' is 1.

在一特定態樣中，本發明係關於一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(V)， 其中方括號中之結構表示該血清型38醣之重複單元，且其中n表示重複單元之數目。 In a specific embodiment, the present invention relates to a serotype 38 saccharide conjugate, which comprises a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and has the general formula (V), Wherein the structure in square brackets represents the repeating unit of the serotype 38 carbohydrate, and wherein n represents the number of repeating units.

式(IV)、(V)為本發明之醣結合物的示意性表示。不應理解為醣之每一重複單元(方括號中之結構)處均存在連接。實情為，大部分醣重複單元保持未經修飾，且少數醣重複單元處存在載體蛋白與醣之共價連接。另外，個別載體蛋白(CP)分子可連接至多於一個醣分子，且個體醣分子可連接至多於一個個別載體蛋白(CP)分子。方括號中之結構表示血清型38醣之重複單元。Formulas (IV) and (V) are schematic representations of the carbohydrate conjugates of the present invention. It should not be understood that there is a connection at every repeat unit of the carbohydrate (structure in square brackets). In fact, most of the carbohydrate repeat units remain unmodified, and a few carbohydrate repeat units have a covalent connection between the carrier protein and the carbohydrate. In addition, an individual carrier protein (CP) molecule can be linked to more than one carbohydrate molecule, and an individual carbohydrate molecule can be linked to more than one individual carrier protein (CP) molecule. The structure in square brackets represents the repeat unit of serotype 38 carbohydrate.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(IV)，其中X為CH 2(CH 2) n '，其中n'係選自1至10，且其中X'為CH 2O(CH 2) n ''CH 2C=O，其中n''係選自0至10。在一實施例中，n'係選自1至5且n''係選自0至10。在一實施例中，n'係選自1至5且n''係選自0至5。在一實施例中，n'係選自1至3且n''係選自0至3。在一實施例中，n'係選自1至2且n''係選自0至2。在一特定實施例中，n'為1且n''為0。在另一實施例中，n'為2且n''為0。在又一實施例中，n'為3且n''為0。在又另一實施例中，n'為4且n''為0。在又另一實施例中，n'為5且n''為0。在又另一實施例中，n'為6且n''為0。在一特定實施例中，n'為1且n''為1。在另一實施例中，n'為2且n''為1。在又一實施例中，n'為3且n''為1。在又另一實施例中，n'為4且n''為1。在又另一實施例中，n'為5且n''為1。在又另一實施例中，n'為6且n''為1。在一特定實施例中，n'為1且n''為2。在另一實施例中，n'為2且n''為2。在又一實施例中，n'為3且n''為2。在又另一實施例中，n'為4且n''為2。在又另一實施例中，n'為5且n''為2。在又另一實施例中，n'為6且n''為2。在一特定實施例中，n'為1且n''為3。在另一實施例中，n'為2且n''為3。在又一實施例中，n'為3且n''為3。在又另一實施例中，n'為4且n''為3。在又另一實施例中，n'為5且n''為3。在又另一實施例中，n'為6且n''為3。在一特定實施例中，n'為1且n''為4。在另一實施例中，n'為2且n''為4。在又一實施例中，n'為3且n''為4。在又另一實施例中，n'為4且n''為4。在又另一實施例中，n'為5且n''為4。在又另一實施例中，n'為6且n''為4。在一特定實施例中，n'為1且n''為5。在另一實施例中，n'為2且n''為5。在又一實施例中，n'為3且n''為5。在又另一實施例中，n'為4且n''為5。在又另一實施例中，n'為5且n''為5。在又另一實施例中，n'為6且n''為5。在一特定實施例中，n'為1且n''為6。在另一實施例中，n'為2且n''為6。在又一實施例中，n'為3且n''為6。在又另一實施例中，n'為4且n''為6。在又另一實施例中，n'為5且n''為6。在又另一實施例中，n'為6且n''為6。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (IV), wherein X is CH 2 (CH 2 ) n ' , wherein n' is selected from 1 to 10, and wherein X' is CH 2 O (CH 2 ) n '' CH 2 C = O, wherein n'' is selected from 0 to 10. In one embodiment, n' is selected from 1 to 5 and n'' is selected from 0 to 10. In one embodiment, n' is selected from 1 to 5 and n'' is selected from 0 to 5. In one embodiment, n' is selected from 1 to 3 and n'' is selected from 0 to 3. In one embodiment, n' is selected from 1 to 2 and n'' is selected from 0 to 2. In a specific embodiment, n' is 1 and n'' is 0. In another embodiment, n' is 2 and n'' is 0. In yet another embodiment, n' is 3 and n'' is 0. In yet another embodiment, n' is 4 and n'' is 0. In yet another embodiment, n' is 5 and n'' is 0. In yet another embodiment, n' is 6 and n'' is 0. In a specific embodiment, n' is 1 and n'' is 1. In another embodiment, n' is 2 and n'' is 1. In yet another embodiment, n' is 3 and n'' is 1. In yet another embodiment, n' is 4 and n'' is 1. In yet another embodiment, n' is 5 and n'' is 1. In yet another embodiment, n' is 6 and n'' is 1. In a specific embodiment, n' is 1 and n'' is 2. In another embodiment, n' is 2 and n'' is 2. In yet another embodiment, n' is 3 and n'' is 2. In yet another embodiment, n' is 4 and n'' is 2. In yet another embodiment, n' is 5 and n'' is 2. In yet another embodiment, n' is 6 and n'' is 2. In a specific embodiment, n' is 1 and n'' is 3. In another embodiment, n' is 2 and n'' is 3. In yet another embodiment, n' is 3 and n'' is 3. In yet another embodiment, n' is 4 and n'' is 3. In yet another embodiment, n' is 5 and n'' is 3. In yet another embodiment, n' is 6 and n'' is 3. In a specific embodiment, n' is 1 and n'' is 4. In another embodiment, n' is 2 and n'' is 4. In yet another embodiment, n' is 3 and n'' is 4. In yet another embodiment, n' is 4 and n'' is 4. In yet another embodiment, n' is 5 and n'' is 4. In yet another embodiment, n' is 6 and n'' is 4. In a specific embodiment, n' is 1 and n'' is 5. In another embodiment, n' is 2 and n'' is 5. In yet another embodiment, n' is 3 and n'' is 5. In yet another embodiment, n' is 4 and n'' is 5. In yet another embodiment, n' is 5 and n'' is 5. In yet another embodiment, n' is 6 and n'' is 5. In a specific embodiment, n' is 1 and n'' is 6. In another embodiment, n' is 2 and n'' is 6. In yet another embodiment, n' is 3 and n'' is 6. In yet another embodiment, n' is 4 and n" is 6. In yet another embodiment, n' is 5 and n" is 6. In yet another embodiment, n' is 6 and n" is 6.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(IV)，其中X為CH 2(CH 2) n '，其中n'係選自1至10，且其中X'為CH 2O(CH 2CH 2O) m '(CH 2) n ''CH 2C=O，其中n''係選自0至10且m'係選自0至4。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (IV), wherein X is CH 2 (CH 2 ) n ′ , wherein n′ is selected from 1 to 10, and wherein X′ is CH 2 O(CH 2 CH 2 O) m ′ (CH 2 ) n ′′ CH 2 C═O, wherein n′′ is selected from 0 to 10 and m′ is selected from 0 to 4.

在一實施例中，n'係選自1至5，m'係選自0至4且n''係選自0至10。在一實施例中，n'係選自1至5，m'係選自0至4且n''係選自0至5。在一實施例中，n'係選自1至3，m'係選自0至2且n''係選自0至3。在一實施例中，n'係選自1至2，m'係選自0至2且n''係選自0至1。In one embodiment, n' is selected from 1 to 5, m' is selected from 0 to 4 and n'' is selected from 0 to 10. In one embodiment, n' is selected from 1 to 5, m' is selected from 0 to 4 and n'' is selected from 0 to 5. In one embodiment, n' is selected from 1 to 3, m' is selected from 0 to 2 and n'' is selected from 0 to 3. In one embodiment, n' is selected from 1 to 2, m' is selected from 0 to 2 and n'' is selected from 0 to 1.

在一特定實施例中，n'為1，m'為0且n''為0。在另一實施例中，n'為1，m'為1且n''為0。在另一實施例中，n'為1，m'為2且n''為0。在另一實施例中，n'為1，m'為3且n''為0。In a particular embodiment, n' is 1, m' is 0 and n'' is 0. In another embodiment, n' is 1, m' is 1 and n'' is 0. In another embodiment, n' is 1, m' is 2 and n'' is 0. In another embodiment, n' is 1, m' is 3 and n'' is 0.

在另一實施例中，n'為2，m'為0且n''為0。在另一實施例中，n'為2，m'為1且n''為0。在另一實施例中，n'為2，m'為2且n''為0。在另一實施例中，n'為2，m'為3且n''為0。In another embodiment, n' is 2, m' is 0 and n'' is 0. In another embodiment, n' is 2, m' is 1 and n'' is 0. In another embodiment, n' is 2, m' is 2 and n'' is 0. In another embodiment, n' is 2, m' is 3 and n'' is 0.

在又一實施例中，n'為3，m'為0且n''為0。在又一實施例中，n'為3，m'為1且n''為0。在又一實施例中，n'為3，m'為2且n''為0。在又一實施例中，n'為3，m'為3且n''為0。In yet another embodiment, n' is 3, m' is 0 and n'' is 0. In yet another embodiment, n' is 3, m' is 1 and n'' is 0. In yet another embodiment, n' is 3, m' is 2 and n'' is 0. In yet another embodiment, n' is 3, m' is 3 and n'' is 0.

在又另一實施例中，n'為4，m'為0且n''為0。在又另一實施例中，n'為4，m'為1且n''為0。在又另一實施例中，n'為4，m'為2且n''為0。在又另一實施例中，n'為4，m'為3且n''為0。In yet another embodiment, n' is 4, m' is 0 and n'' is 0. In yet another embodiment, n' is 4, m' is 1 and n'' is 0. In yet another embodiment, n' is 4, m' is 2 and n'' is 0. In yet another embodiment, n' is 4, m' is 3 and n'' is 0.

在又另一實施例中，n'為5，m'為0且n''為0。在又另一實施例中，n'為5，m'為1且n''為0。在又另一實施例中，n'為5，m'為2且n''為0。在又另一實施例中，n'為5，m'為3且n''為0。In yet another embodiment, n' is 5, m' is 0 and n'' is 0. In yet another embodiment, n' is 5, m' is 1 and n'' is 0. In yet another embodiment, n' is 5, m' is 2 and n'' is 0. In yet another embodiment, n' is 5, m' is 3 and n'' is 0.

在一特定實施例中，n'為1，m'為0且n''為1。在一特定實施例中，n'為1，m'為1且n''為1。在一特定實施例中，n'為1，m'為2且n''為1。在一特定實施例中，n'為1，m'為3且n''為1。In a specific embodiment, n' is 1, m' is 0 and n'' is 1. In a specific embodiment, n' is 1, m' is 1 and n'' is 1. In a specific embodiment, n' is 1, m' is 2 and n'' is 1. In a specific embodiment, n' is 1, m' is 3 and n'' is 1.

在另一實施例中，n'為2，m'為0且n''為1。在另一實施例中，n'為2，m'為1且n''為1。在另一實施例中，n'為2，m'為2且n''為1。在另一實施例中，n'為2，m'為3且n''為1。In another embodiment, n' is 2, m' is 0 and n'' is 1. In another embodiment, n' is 2, m' is 1 and n'' is 1. In another embodiment, n' is 2, m' is 2 and n'' is 1. In another embodiment, n' is 2, m' is 3 and n'' is 1.

在又一實施例中，n'為3，m'為0且n''為1。在又一實施例中，n'為3，m'為1且n''為1。在又一實施例中，n'為3，m'為2且n''為1。在又一實施例中，n'為3，m'為3且n''為1。In yet another embodiment, n' is 3, m' is 0 and n'' is 1. In yet another embodiment, n' is 3, m' is 1 and n'' is 1. In yet another embodiment, n' is 3, m' is 2 and n'' is 1. In yet another embodiment, n' is 3, m' is 3 and n'' is 1.

在又另一實施例中，n'為4，m'為0且n''為1。在又另一實施例中，n'為4，m'為1且n''為1。在又另一實施例中，n'為4，m'為2且n''為1。在又另一實施例中，n'為4，m'為3且n''為1。In yet another embodiment, n' is 4, m' is 0 and n" is 1. In yet another embodiment, n' is 4, m' is 1 and n" is 1. In yet another embodiment, n' is 4, m' is 2 and n" is 1. In yet another embodiment, n' is 4, m' is 3 and n" is 1.

在又另一實施例中，n'為5，m'為0且n''為1。在又另一實施例中，n'為5，m'為1且n''為1。在又另一實施例中，n'為5，m'為2且n''為1。在又另一實施例中，n'為5，m'為3且n''為1。In yet another embodiment, n' is 5, m' is 0 and n'' is 1. In yet another embodiment, n' is 5, m' is 1 and n'' is 1. In yet another embodiment, n' is 5, m' is 2 and n'' is 1. In yet another embodiment, n' is 5, m' is 3 and n'' is 1.

在一特定實施例中，n'為1，m'為0且n''為2。在一特定實施例中，n'為1，m'為1且n''為2。在一特定實施例中，n'為1，m'為2且n''為2。在一特定實施例中，n'為1，m'為3且n''為2。In a specific embodiment, n' is 1, m' is 0 and n'' is 2. In a specific embodiment, n' is 1, m' is 1 and n'' is 2. In a specific embodiment, n' is 1, m' is 2 and n'' is 2. In a specific embodiment, n' is 1, m' is 3 and n'' is 2.

在另一實施例中，n'為2，m'為0且n''為2。在另一實施例中，n'為2，m'為1且n''為2。在另一實施例中，n'為2，m'為2且n''為2。在另一實施例中，n'為2，m'為3且n''為2。In another embodiment, n' is 2, m' is 0 and n'' is 2. In another embodiment, n' is 2, m' is 1 and n'' is 2. In another embodiment, n' is 2, m' is 2 and n'' is 2. In another embodiment, n' is 2, m' is 3 and n'' is 2.

在又一實施例中，n'為3，m'為0且n''為2。在又一實施例中，n'為3，m'為1且n''為2。在又一實施例中，n'為3，m'為2且n''為2。在又一實施例中，n'為3，m'為3且n''為2。In yet another embodiment, n' is 3, m' is 0 and n'' is 2. In yet another embodiment, n' is 3, m' is 1 and n'' is 2. In yet another embodiment, n' is 3, m' is 2 and n'' is 2. In yet another embodiment, n' is 3, m' is 3 and n'' is 2.

在又另一實施例中，n'為4，m'為0且n''為2。在又另一實施例中，n'為4，m'為1且n''為2。在又另一實施例中，n'為4，m'為2且n''為2。在又另一實施例中，n'為4，m'為3且n''為2。In yet another embodiment, n' is 4, m' is 0 and n" is 2. In yet another embodiment, n' is 4, m' is 1 and n" is 2. In yet another embodiment, n' is 4, m' is 2 and n" is 2. In yet another embodiment, n' is 4, m' is 3 and n" is 2.

在又另一實施例中，n'為5，m'為0且n''為2。在又另一實施例中，n'為5，m'為1且n''為2。在又另一實施例中，n'為5，m'為2且n''為2。在又另一實施例中，n'為5，m'為3且n''為2。In yet another embodiment, n' is 5, m' is 0 and n'' is 2. In yet another embodiment, n' is 5, m' is 1 and n'' is 2. In yet another embodiment, n' is 5, m' is 2 and n'' is 2. In yet another embodiment, n' is 5, m' is 3 and n'' is 2.

在一特定實施例中，n'為1，m'為0且n''為3。在一特定實施例中，n'為1，m'為1且n''為3。在一特定實施例中，n'為1，m'為2且n''為3。在一特定實施例中，n'為1，m'為3且n''為3。In a specific embodiment, n' is 1, m' is 0 and n'' is 3. In a specific embodiment, n' is 1, m' is 1 and n'' is 3. In a specific embodiment, n' is 1, m' is 2 and n'' is 3. In a specific embodiment, n' is 1, m' is 3 and n'' is 3.

在另一實施例中，n'為2，m'為0且n''為3。在另一實施例中，n'為2，m'為1且n''為3。在另一實施例中，n'為2，m'為2且n''為3。在另一實施例中，n'為2，m'為3且n''為3。In another embodiment, n' is 2, m' is 0 and n'' is 3. In another embodiment, n' is 2, m' is 1 and n'' is 3. In another embodiment, n' is 2, m' is 2 and n'' is 3. In another embodiment, n' is 2, m' is 3 and n'' is 3.

在又一實施例中，n'為3，m'為0且n''為3。在又一實施例中，n'為3，m'為1且n''為3。在又一實施例中，n'為3，m'為2且n''為3。在又一實施例中，n'為3，m'為3且n''為3。In yet another embodiment, n' is 3, m' is 0 and n'' is 3. In yet another embodiment, n' is 3, m' is 1 and n'' is 3. In yet another embodiment, n' is 3, m' is 2 and n'' is 3. In yet another embodiment, n' is 3, m' is 3 and n'' is 3.

在又另一實施例中，n'為4，m'為0且n''為3。在又另一實施例中，n'為4，m'為1且n''為3。在又另一實施例中，n'為4，m'為2且n''為3。在又另一實施例中，n'為4，m'為3且n''為3。In yet another embodiment, n' is 4, m' is 0 and n'' is 3. In yet another embodiment, n' is 4, m' is 1 and n'' is 3. In yet another embodiment, n' is 4, m' is 2 and n'' is 3. In yet another embodiment, n' is 4, m' is 3 and n'' is 3.

在又另一實施例中，n'為5，m'為0且n''為3。在又另一實施例中，n'為5，m'為1且n''為3。在又另一實施例中，n'為5，m'為2且n''為3。在又另一實施例中，n'為5，m'為3且n''為3。In yet another embodiment, n' is 5, m' is 0 and n'' is 3. In yet another embodiment, n' is 5, m' is 1 and n'' is 3. In yet another embodiment, n' is 5, m' is 2 and n'' is 3. In yet another embodiment, n' is 5, m' is 3 and n'' is 3.

在一特定實施例中，n'為1，m'為0且n''為4。在一特定實施例中，n'為1，m'為1且n''為4。在一特定實施例中，n'為1，m'為2且n''為4。在一特定實施例中，n'為1，m'為3且n''為4。In a specific embodiment, n' is 1, m' is 0 and n'' is 4. In a specific embodiment, n' is 1, m' is 1 and n'' is 4. In a specific embodiment, n' is 1, m' is 2 and n'' is 4. In a specific embodiment, n' is 1, m' is 3 and n'' is 4.

在另一實施例中，n'為2，m'為0且n''為4。在另一實施例中，n'為2，m'為1且n''為4。在另一實施例中，n'為2，m'為2且n''為4。在另一實施例中，n'為2，m'為3且n''為4。In another embodiment, n' is 2, m' is 0 and n'' is 4. In another embodiment, n' is 2, m' is 1 and n'' is 4. In another embodiment, n' is 2, m' is 2 and n'' is 4. In another embodiment, n' is 2, m' is 3 and n'' is 4.

在又一實施例中，n'為3，m'為0且n''為4。在又一實施例中，n'為3，m'為1且n''為4。在又一實施例中，n'為3，m'為2且n''為4。在又一實施例中，n'為3，m'為3且n''為4。In yet another embodiment, n' is 3, m' is 0 and n'' is 4. In yet another embodiment, n' is 3, m' is 1 and n'' is 4. In yet another embodiment, n' is 3, m' is 2 and n'' is 4. In yet another embodiment, n' is 3, m' is 3 and n'' is 4.

在又另一實施例中，n'為4，m'為0且n''為4。在又另一實施例中，n'為4，m'為1且n''為4。在又另一實施例中，n'為4，m'為2且n''為4。在又另一實施例中，n'為4，m'為3且n''為4。In yet another embodiment, n' is 4, m' is 0 and n'' is 4. In yet another embodiment, n' is 4, m' is 1 and n'' is 4. In yet another embodiment, n' is 4, m' is 2 and n'' is 4. In yet another embodiment, n' is 4, m' is 3 and n'' is 4.

在又另一實施例中，n'為5，m'為0且n''為4。在又另一實施例中，n'為5，m'為1且n''為4。在又另一實施例中，n'為5，m'為2且n''為4。在又另一實施例中，n'為5，m'為3且n''為4。In yet another embodiment, n' is 5, m' is 0 and n'' is 4. In yet another embodiment, n' is 5, m' is 1 and n'' is 4. In yet another embodiment, n' is 5, m' is 2 and n'' is 4. In yet another embodiment, n' is 5, m' is 3 and n'' is 4.

在一特定實施例中，n'為1，m'為0且n''為5。在一特定實施例中，n'為1，m'為1且n''為5。在一特定實施例中，n'為1，m'為2且n''為5。在一特定實施例中，n'為1，m'為3且n''為5。In a specific embodiment, n' is 1, m' is 0 and n'' is 5. In a specific embodiment, n' is 1, m' is 1 and n'' is 5. In a specific embodiment, n' is 1, m' is 2 and n'' is 5. In a specific embodiment, n' is 1, m' is 3 and n'' is 5.

在另一實施例中，n'為2，m'為0且n''為5。在另一實施例中，n'為2，m'為1且n''為5。在另一實施例中，n'為2，m'為2且n''為5。在另一實施例中，n'為2，m'為3且n''為5。In another embodiment, n' is 2, m' is 0 and n'' is 5. In another embodiment, n' is 2, m' is 1 and n'' is 5. In another embodiment, n' is 2, m' is 2 and n'' is 5. In another embodiment, n' is 2, m' is 3 and n'' is 5.

在又一實施例中，n'為3，m'為0且n''為5。在又一實施例中，n'為3，m'為1且n''為5。在又一實施例中，n'為3，m'為2且n''為5。在又一實施例中，n'為3，m'為3且n''為5。In yet another embodiment, n' is 3, m' is 0 and n'' is 5. In yet another embodiment, n' is 3, m' is 1 and n'' is 5. In yet another embodiment, n' is 3, m' is 2 and n'' is 5. In yet another embodiment, n' is 3, m' is 3 and n'' is 5.

在又另一實施例中，n'為4，m'為0且n''為5。在又另一實施例中，n'為4，m'為1且n''為5。在又另一實施例中，n'為4，m'為2且n''為5。在又另一實施例中，n'為4，m'為3且n''為5。In yet another embodiment, n' is 4, m' is 0 and n'' is 5. In yet another embodiment, n' is 4, m' is 1 and n'' is 5. In yet another embodiment, n' is 4, m' is 2 and n'' is 5. In yet another embodiment, n' is 4, m' is 3 and n'' is 5.

在又另一實施例中，n'為5，m'為0且n''為5。在又另一實施例中，n'為5，m'為1且n''為5。在又另一實施例中，n'為5，m'為2且n''為5。在又另一實施例中，n'為5，m'為3且n''為5。In yet another embodiment, n' is 5, m' is 0 and n'' is 5. In yet another embodiment, n' is 5, m' is 1 and n'' is 5. In yet another embodiment, n' is 5, m' is 2 and n'' is 5. In yet another embodiment, n' is 5, m' is 3 and n'' is 5.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(IV)，其中X為(CH 2CH 2O) mCH 2CH 2，其中m係選自1至4，且其中X'為CH 2O(CH 2) n ''CH 2C=O，其中n''係選自0至10。在一實施例中，m係選自1至3且n''係選自0至10。在一實施例中，m係選自1至3且n''係選自0至5。在一實施例中，m係選自1至2且n''係選自0至3。在一實施例中，m係選自1至2且n''係選自0至2。在一特定實施例中，m為1且n''為0。在另一實施例中，m為2且n''為0。在又一實施例中，m為3且n''為0。在又另一實施例中，m為4且n''為0。在一特定實施例中，m為1且n''為1。在另一實施例中，m為2且n''為1。在又一實施例中，m為3且n''為1。在又另一實施例中，m為4且n''為1。在一特定實施例中，m為1且n''為2。在另一實施例中，m為2且n''為2。在又一實施例中，m為3且n''為2。在又另一實施例中，m為4且n''為2。在一特定實施例中，m為1且n''為3。在另一實施例中，m為2且n''為3。在又一實施例中，m為3且n''為3。在又另一實施例中，m為4且n''為3。在一特定實施例中，m為1且n''為4。在另一實施例中，m為2且n''為4。在又一實施例中，m為3且n''為4。在又另一實施例中，m為4且n''為4。在一特定實施例中，m為1且n''為5。在另一實施例中，m為2且n''為5。在又一實施例中，m為3且n''為5。在又另一實施例中，m為4且n''為5。在一特定實施例中，m為1且n''為6。在另一實施例中，m為2且n''為6。在又一實施例中，m為3且n''為6。在又另一實施例中，m為4且n''為6。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (IV), wherein X is (CH 2 CH 2 O) m CH 2 CH 2 , wherein m is selected from 1 to 4, and wherein X' is CH 2 O(CH 2 ) n '' CH 2 C=O, wherein n '' is selected from 0 to 10. In one embodiment, m is selected from 1 to 3 and n '' is selected from 0 to 10. In one embodiment, m is selected from 1 to 3 and n '' is selected from 0 to 5. In one embodiment, m is selected from 1 to 2 and n '' is selected from 0 to 3. In one embodiment, m is selected from 1 to 2 and n '' is selected from 0 to 2. In a specific embodiment, m is 1 and n '' is 0. In another embodiment, m is 2 and n'' is 0. In yet another embodiment, m is 3 and n'' is 0. In yet another embodiment, m is 4 and n'' is 0. In a specific embodiment, m is 1 and n'' is 1. In another embodiment, m is 2 and n'' is 1. In yet another embodiment, m is 3 and n'' is 1. In yet another embodiment, m is 4 and n'' is 1. In a specific embodiment, m is 1 and n'' is 2. In another embodiment, m is 2 and n'' is 2. In yet another embodiment, m is 3 and n'' is 2. In yet another embodiment, m is 4 and n'' is 2. In a specific embodiment, m is 1 and n'' is 3. In another embodiment, m is 2 and n'' is 3. In yet another embodiment, m is 3 and n'' is 3. In yet another embodiment, m is 4 and n'' is 3. In a specific embodiment, m is 1 and n'' is 4. In another embodiment, m is 2 and n'' is 4. In yet another embodiment, m is 3 and n'' is 4. In yet another embodiment, m is 4 and n'' is 4. In a specific embodiment, m is 1 and n'' is 5. In another embodiment, m is 2 and n'' is 5. In yet another embodiment, m is 3 and n'' is 5. In yet another embodiment, m is 4 and n'' is 5. In a specific embodiment, m is 1 and n'' is 6. In another embodiment, m is 2 and n'' is 6. In yet another embodiment, m is 3 and n'' is 6. In yet another embodiment, m is 4 and n″ is 6.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(IV)，其中X為(CH 2CH 2O) mCH 2CH 2，其中m係選自1至4，且其中X'為CH 2O(CH 2CH 2O) m '(CH 2) n ''CH 2C=O，其中n''係選自0至10且m'係選自0至4。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (IV), wherein X is (CH 2 CH 2 O) m CH 2 CH 2 , wherein m is selected from 1 to 4, and wherein X' is CH 2 O(CH 2 CH 2 O) m ' (CH 2 ) n '' CH 2 C=O, wherein n'' is selected from 0 to 10 and m' is selected from 0 to 4.

在一實施例中，m係選自1至3，m'係選自0至4且n''係選自0至10。在一實施例中，m係選自1至2，m'係選自0至4且n''係選自0至5。在一實施例中，m係選自1至2，m'係選自0至2且n''係選自0至3。在一實施例中，m係選自1至2，m'係選自0至2且n''係選自0至1。In one embodiment, m is selected from 1 to 3, m' is selected from 0 to 4 and n'' is selected from 0 to 10. In one embodiment, m is selected from 1 to 2, m' is selected from 0 to 4 and n'' is selected from 0 to 5. In one embodiment, m is selected from 1 to 2, m' is selected from 0 to 2 and n'' is selected from 0 to 3. In one embodiment, m is selected from 1 to 2, m' is selected from 0 to 2 and n'' is selected from 0 to 1.

在一特定實施例中，m為1，m'為0且n''為0。在另一實施例中，m為1，m'為1且n''為0。在另一實施例中，m為1，m'為2且n''為0。在另一實施例中，m為1，m'為3且n''為0。In a particular embodiment, m is 1, m' is 0 and n'' is 0. In another embodiment, m is 1, m' is 1 and n'' is 0. In another embodiment, m is 1, m' is 2 and n'' is 0. In another embodiment, m is 1, m' is 3 and n'' is 0.

在另一實施例中，m為2，m'為0且n''為0。在另一實施例中，m為2，m'為1且n''為0。在另一實施例中，m為2，m'為2且n''為0。在另一實施例中，m為2，m'為3且n''為0。In another embodiment, m is 2, m' is 0 and n'' is 0. In another embodiment, m is 2, m' is 1 and n'' is 0. In another embodiment, m is 2, m' is 2 and n'' is 0. In another embodiment, m is 2, m' is 3 and n'' is 0.

在又一實施例中，m為3，m'為0且n''為0。在又一實施例中，m為3，m'為1且n''為0。在又一實施例中，m為3，m'為2且n''為0。在又一實施例中，m為3，m'為3且n''為0。In yet another embodiment, m is 3, m' is 0 and n'' is 0. In yet another embodiment, m is 3, m' is 1 and n'' is 0. In yet another embodiment, m is 3, m' is 2 and n'' is 0. In yet another embodiment, m is 3, m' is 3 and n'' is 0.

在又另一實施例中，m為4，m'為0且n''為0。在又另一實施例中，m為4，m'為1且n''為0。在又另一實施例中，m為4，m'為2且n''為0。在又另一實施例中，m為4，m'為3且n''為0。In yet another embodiment, m is 4, m' is 0 and n" is 0. In yet another embodiment, m is 4, m' is 1 and n" is 0. In yet another embodiment, m is 4, m' is 2 and n" is 0. In yet another embodiment, m is 4, m' is 3 and n" is 0.

在一特定實施例中，m為1，m'為0且n''為1。在一特定實施例中，m為1，m'為1且n''為1。在一特定實施例中，m為1，m'為2且n''為1。在一特定實施例中，m為1，m'為3且n''為1。In a specific embodiment, m is 1, m' is 0 and n'' is 1. In a specific embodiment, m is 1, m' is 1 and n'' is 1. In a specific embodiment, m is 1, m' is 2 and n'' is 1. In a specific embodiment, m is 1, m' is 3 and n'' is 1.

在另一實施例中，m為2，m'為0且n''為1。在另一實施例中，m為2，m'為1且n''為1。在另一實施例中，m為2，m'為2且n''為1。在另一實施例中，m為2，m'為3且n''為1。In another embodiment, m is 2, m' is 0 and n'' is 1. In another embodiment, m is 2, m' is 1 and n'' is 1. In another embodiment, m is 2, m' is 2 and n'' is 1. In another embodiment, m is 2, m' is 3 and n'' is 1.

在又一實施例中，m為3，m'為0且n''為1。在又一實施例中，m為3，m'為1且n''為1。在又一實施例中，m為3，m'為2且n''為1。在又一實施例中，m為3，m'為3且n''為1。In yet another embodiment, m is 3, m' is 0 and n'' is 1. In yet another embodiment, m is 3, m' is 1 and n'' is 1. In yet another embodiment, m is 3, m' is 2 and n'' is 1. In yet another embodiment, m is 3, m' is 3 and n'' is 1.

在又另一實施例中，m為4，m'為0且n''為1。在又另一實施例中，m為4，m'為1且n''為1。在又另一實施例中，m為4，m'為2且n''為1。在又另一實施例中，m為4，m'為3且n''為1。In yet another embodiment, m is 4, m' is 0 and n'' is 1. In yet another embodiment, m is 4, m' is 1 and n'' is 1. In yet another embodiment, m is 4, m' is 2 and n'' is 1. In yet another embodiment, m is 4, m' is 3 and n'' is 1.

在一特定實施例中，m為1，m'為0且n''為2。在一特定實施例中，m為1，m'為1且n''為2。在一特定實施例中，m為1，m'為2且n''為2。在一特定實施例中，m為1，m'為3且n''為2。In a specific embodiment, m is 1, m' is 0 and n'' is 2. In a specific embodiment, m is 1, m' is 1 and n'' is 2. In a specific embodiment, m is 1, m' is 2 and n'' is 2. In a specific embodiment, m is 1, m' is 3 and n'' is 2.

在另一實施例中，m為2，m'為0且n''為2。在另一實施例中，m為2，m'為1且n''為2。在另一實施例中，m為2，m'為2且n''為2。在另一實施例中，m為2，m'為3且n''為2。In another embodiment, m is 2, m' is 0 and n'' is 2. In another embodiment, m is 2, m' is 1 and n'' is 2. In another embodiment, m is 2, m' is 2 and n'' is 2. In another embodiment, m is 2, m' is 3 and n'' is 2.

在又一實施例中，m為3，m'為0且n''為2。在又一實施例中，m為3，m'為1且n''為2。在又一實施例中，m為3，m'為2且n''為2。在又一實施例中，m為3，m'為3且n''為2。In yet another embodiment, m is 3, m' is 0 and n'' is 2. In yet another embodiment, m is 3, m' is 1 and n'' is 2. In yet another embodiment, m is 3, m' is 2 and n'' is 2. In yet another embodiment, m is 3, m' is 3 and n'' is 2.

在又另一實施例中，m為4，m'為0且n''為2。在又另一實施例中，m為4，m'為1且n''為2。在又另一實施例中，m為4，m'為2且n''為2。在又另一實施例中，m為4，m'為3且n''為2。In yet another embodiment, m is 4, m' is 0 and n'' is 2. In yet another embodiment, m is 4, m' is 1 and n'' is 2. In yet another embodiment, m is 4, m' is 2 and n'' is 2. In yet another embodiment, m is 4, m' is 3 and n'' is 2.

在一特定實施例中，m為1，m'為0且n''為3。在一特定實施例中，m為1，m'為1且n''為3。在一特定實施例中，m為1，m'為2且n''為3。在一特定實施例中，m為1，m'為3且n''為3。In a specific embodiment, m is 1, m' is 0 and n'' is 3. In a specific embodiment, m is 1, m' is 1 and n'' is 3. In a specific embodiment, m is 1, m' is 2 and n'' is 3. In a specific embodiment, m is 1, m' is 3 and n'' is 3.

在另一實施例中，m為2，m'為0且n''為3。在另一實施例中，m為2，m'為1且n''為3。在另一實施例中，m為2，m'為2且n''為3。在另一實施例中，m為2，m'為3且n''為3。In another embodiment, m is 2, m' is 0 and n'' is 3. In another embodiment, m is 2, m' is 1 and n'' is 3. In another embodiment, m is 2, m' is 2 and n'' is 3. In another embodiment, m is 2, m' is 3 and n'' is 3.

在又一實施例中，m為3，m'為0且n''為3。在又一實施例中，m為3，m'為1且n''為3。在又一實施例中，m為3，m'為2且n''為3。在又一實施例中，m為3，m'為3且n''為3。In yet another embodiment, m is 3, m' is 0 and n'' is 3. In yet another embodiment, m is 3, m' is 1 and n'' is 3. In yet another embodiment, m is 3, m' is 2 and n'' is 3. In yet another embodiment, m is 3, m' is 3 and n'' is 3.

在又另一實施例中，m為4，m'為0且n''為3。在又另一實施例中，m為4，m'為1且n''為3。在又另一實施例中，m為4，m'為2且n''為3。在又另一實施例中，m為4，m'為3且n''為3。In yet another embodiment, m is 4, m' is 0 and n'' is 3. In yet another embodiment, m is 4, m' is 1 and n'' is 3. In yet another embodiment, m is 4, m' is 2 and n'' is 3. In yet another embodiment, m is 4, m' is 3 and n'' is 3.

在一特定實施例中，m為1，m'為0且n''為4。在一特定實施例中，m為1，m'為1且n''為4。在一特定實施例中，m為1，m'為2且n''為4。在一特定實施例中，m為1，m'為3且n''為4。In a specific embodiment, m is 1, m' is 0 and n'' is 4. In a specific embodiment, m is 1, m' is 1 and n'' is 4. In a specific embodiment, m is 1, m' is 2 and n'' is 4. In a specific embodiment, m is 1, m' is 3 and n'' is 4.

在另一實施例中，m為2，m'為0且n''為4。在另一實施例中，m為2，m'為1且n''為4。在另一實施例中，m為2，m'為2且n''為4。在另一實施例中，m為2，m'為3且n''為4。In another embodiment, m is 2, m' is 0 and n'' is 4. In another embodiment, m is 2, m' is 1 and n'' is 4. In another embodiment, m is 2, m' is 2 and n'' is 4. In another embodiment, m is 2, m' is 3 and n'' is 4.

在又一實施例中，m為3，m'為0且n''為4。在又一實施例中，m為3，m'為1且n''為4。在又一實施例中，m為3，m'為2且n''為4。在又一實施例中，m為3，m'為3且n''為4。In yet another embodiment, m is 3, m' is 0 and n'' is 4. In yet another embodiment, m is 3, m' is 1 and n'' is 4. In yet another embodiment, m is 3, m' is 2 and n'' is 4. In yet another embodiment, m is 3, m' is 3 and n'' is 4.

在又另一實施例中，m為4，m'為0且n''為4。在又另一實施例中，m為4，m'為1且n''為4。在又另一實施例中，m為4，m'為2且n''為4。在又另一實施例中，m為4，m'為3且n''為4。In yet another embodiment, m is 4, m' is 0 and n'' is 4. In yet another embodiment, m is 4, m' is 1 and n'' is 4. In yet another embodiment, m is 4, m' is 2 and n'' is 4. In yet another embodiment, m is 4, m' is 3 and n'' is 4.

在一特定實施例中，m為1，m'為0且n''為5。在一特定實施例中，m為1，m'為1且n''為5。在一特定實施例中，m為1，m'為2且n''為5。在一特定實施例中，m為1，m'為3且n''為5。In a specific embodiment, m is 1, m' is 0 and n'' is 5. In a specific embodiment, m is 1, m' is 1 and n'' is 5. In a specific embodiment, m is 1, m' is 2 and n'' is 5. In a specific embodiment, m is 1, m' is 3 and n'' is 5.

在另一實施例中，m為2，m'為0且n''為5。在另一實施例中，m為2，m'為1且n''為5。在另一實施例中，m為2，m'為2且n''為5。在另一實施例中，m為2，m'為3且n''為5。In another embodiment, m is 2, m' is 0 and n'' is 5. In another embodiment, m is 2, m' is 1 and n'' is 5. In another embodiment, m is 2, m' is 2 and n'' is 5. In another embodiment, m is 2, m' is 3 and n'' is 5.

在又一實施例中，m為3，m'為0且n''為5。在又一實施例中，m為3，m'為1且n''為5。在又一實施例中，m為3，m'為2且n''為5。在又一實施例中，m為3，m'為3且n''為5。In yet another embodiment, m is 3, m' is 0 and n'' is 5. In yet another embodiment, m is 3, m' is 1 and n'' is 5. In yet another embodiment, m is 3, m' is 2 and n'' is 5. In yet another embodiment, m is 3, m' is 3 and n'' is 5.

在又另一實施例中，m為4，m'為0且n''為5。在又另一實施例中，m為4，m'為1且n''為5。在又另一實施例中，m為4，m'為2且n''為5。在又另一實施例中，m為4，m'為3且n''為5。In yet another embodiment, m is 4, m' is 0 and n'' is 5. In yet another embodiment, m is 4, m' is 1 and n'' is 5. In yet another embodiment, m is 4, m' is 2 and n'' is 5. In yet another embodiment, m is 4, m' is 3 and n'' is 5.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(IV)，其中X為NHCO(CH 2) n '，其中n'係選自1至10，且其中X'為CH 2O(CH 2) n ''CH 2C=O，其中n''係選自0至10。在一實施例中，n'係選自1至5且n''係選自0至10。在一實施例中，n'係選自1至5且n''係選自0至5。在一實施例中，n'係選自1至3且n''係選自0至3。在一實施例中，n'係選自1至2且n''係選自0至2。在一特定實施例中，n'為1且n''為0。在另一實施例中，n'為2且n''為0。在又一實施例中，n'為3且n''為0。在又另一實施例中，n'為4且n''為0。在又另一實施例中，n'為5且n''為0。在又另一實施例中，n'為6且n''為0。在一特定實施例中，n'為1且n''為1。在另一實施例中，n'為2且n''為1。在又一實施例中，n'為3且n''為1。在又另一實施例中，n'為4且n''為1。在又另一實施例中，n'為5且n''為1。在又另一實施例中，n'為6且n''為1。在一特定實施例中，n'為1且n''為2。在另一實施例中，n'為2且n''為2。在又一實施例中，n'為3且n''為2。在又另一實施例中，n'為4且n''為2。在又另一實施例中，n'為5且n''為2。在又另一實施例中，n'為6且n''為2。在一特定實施例中，n'為1且n''為3。在另一實施例中，n'為2且n''為3。在又一實施例中，n'為3且n''為3。在又另一實施例中，n'為4且n''為3。在又另一實施例中，n'為5且n''為3。在又另一實施例中，n'為6且n''為3。在一特定實施例中，n'為1且n''為4。在另一實施例中，n'為2且n''為4。在又一實施例中，n'為3且n''為4。在又另一實施例中，n'為4且n''為4。在又另一實施例中，n'為5且n''為4。在又另一實施例中，n'為6且n''為4。在一特定實施例中，n'為1且n''為5。在另一實施例中，n'為2且n''為5。在又一實施例中，n'為3且n''為5。在又另一實施例中，n'為4且n''為5。在又另一實施例中，n'為5且n''為5。在又另一實施例中，n'為6且n''為5。在一特定實施例中，n'為1且n''為6。在另一實施例中，n'為2且n''為6。在又一實施例中，n'為3且n''為6。在又另一實施例中，n'為4且n''為6。在又另一實施例中，n'為5且n''為6。在又另一實施例中，n'為6且n''為6。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (IV), wherein X is NHCO(CH 2 ) n ′ , wherein n′ is selected from 1 to 10, and wherein X′ is CH 2 O(CH 2 ) n ′′ CH 2 C═O, wherein n′ is selected from 0 to 10. In one embodiment, n′ is selected from 1 to 5 and n′′ is selected from 0 to 10. In one embodiment, n′ is selected from 1 to 5 and n′′ is selected from 0 to 5. In one embodiment, n′ is selected from 1 to 3 and n′′ is selected from 0 to 3. In one embodiment, n′ is selected from 1 to 2 and n′′ is selected from 0 to 2. In a specific embodiment, n′ is 1 and n′′ is 0. In another embodiment, n' is 2 and n'' is 0. In yet another embodiment, n' is 3 and n'' is 0. In yet another embodiment, n' is 4 and n'' is 0. In yet another embodiment, n' is 5 and n'' is 0. In yet another embodiment, n' is 6 and n'' is 0. In a specific embodiment, n' is 1 and n'' is 1. In another embodiment, n' is 2 and n'' is 1. In yet another embodiment, n' is 3 and n'' is 1. In yet another embodiment, n' is 4 and n'' is 1. In yet another embodiment, n' is 5 and n'' is 1. In yet another embodiment, n' is 6 and n'' is 1. In a specific embodiment, n' is 1 and n'' is 2. In another embodiment, n' is 2 and n'' is 2. In yet another embodiment, n' is 3 and n'' is 2. In yet another embodiment, n' is 4 and n'' is 2. In yet another embodiment, n' is 5 and n'' is 2. In yet another embodiment, n' is 6 and n'' is 2. In a specific embodiment, n' is 1 and n'' is 3. In another embodiment, n' is 2 and n'' is 3. In yet another embodiment, n' is 3 and n'' is 3. In yet another embodiment, n' is 4 and n'' is 3. In yet another embodiment, n' is 5 and n'' is 3. In yet another embodiment, n' is 6 and n'' is 3. In a specific embodiment, n' is 1 and n'' is 4. In another embodiment, n' is 2 and n'' is 4. In yet another embodiment, n' is 3 and n'' is 4. In yet another embodiment, n' is 4 and n'' is 4. In yet another embodiment, n' is 5 and n'' is 4. In yet another embodiment, n' is 6 and n'' is 4. In a specific embodiment, n' is 1 and n'' is 5. In another embodiment, n' is 2 and n'' is 5. In yet another embodiment, n' is 3 and n'' is 5. In yet another embodiment, n' is 4 and n'' is 5. In yet another embodiment, n' is 5 and n'' is 5. In yet another embodiment, n' is 6 and n'' is 5. In a specific embodiment, n' is 1 and n'' is 6. In another embodiment, n' is 2 and n'' is 6. In yet another embodiment, n' is 3 and n'' is 6. In yet another embodiment, n' is 4 and n'' is 6. In yet another embodiment, n' is 5 and n" is 6. In yet another embodiment, n' is 6 and n" is 6.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(IV)，其中X為NHCO(CH 2) n '，其中n'係選自1至10，且其中X'為CH 2O(CH 2CH 2O) m '(CH 2) n ''CH 2C=O，其中n''係選自0至10且m'係選自0至4。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (IV), wherein X is NHCO(CH 2 ) n ′ , wherein n′ is selected from 1 to 10, and wherein X′ is CH 2 O(CH 2 CH 2 O) m ′ (CH 2 ) n ′′ CH 2 C═O, wherein n′′ is selected from 0 to 10 and m′ is selected from 0 to 4.

在一實施例中，n'係選自1至5，m'係選自0至4且n''係選自0至10。在一實施例中，n'係選自1至5，m'係選自0至4且n''係選自0至5。在一實施例中，n'係選自1至3，m'係選自0至2且n''係選自0至3。在一實施例中，n'係選自1至2，m'係選自0至2且n''係選自0至1。In one embodiment, n' is selected from 1 to 5, m' is selected from 0 to 4 and n'' is selected from 0 to 10. In one embodiment, n' is selected from 1 to 5, m' is selected from 0 to 4 and n'' is selected from 0 to 5. In one embodiment, n' is selected from 1 to 3, m' is selected from 0 to 2 and n'' is selected from 0 to 3. In one embodiment, n' is selected from 1 to 2, m' is selected from 0 to 2 and n'' is selected from 0 to 1.

在一特定實施例中，n'為1，m'為0且n''為0。在另一實施例中，n'為1，m'為1且n''為0。在另一實施例中，n'為1，m'為2且n''為0。在另一實施例中，n'為1，m'為3且n''為0。In a particular embodiment, n' is 1, m' is 0 and n'' is 0. In another embodiment, n' is 1, m' is 1 and n'' is 0. In another embodiment, n' is 1, m' is 2 and n'' is 0. In another embodiment, n' is 1, m' is 3 and n'' is 0.

在另一實施例中，n'為2，m'為0且n''為0。在另一實施例中，n'為2，m'為1且n''為0。在另一實施例中，n'為2，m'為2且n''為0。在另一實施例中，n'為2，m'為3且n''為0。In another embodiment, n' is 2, m' is 0 and n'' is 0. In another embodiment, n' is 2, m' is 1 and n'' is 0. In another embodiment, n' is 2, m' is 2 and n'' is 0. In another embodiment, n' is 2, m' is 3 and n'' is 0.

在又一實施例中，n'為3，m'為0且n''為0。在又一實施例中，n'為3，m'為1且n''為0。在又一實施例中，n'為3，m'為2且n''為0。在又一實施例中，n'為3，m'為3且n''為0。In yet another embodiment, n' is 3, m' is 0 and n'' is 0. In yet another embodiment, n' is 3, m' is 1 and n'' is 0. In yet another embodiment, n' is 3, m' is 2 and n'' is 0. In yet another embodiment, n' is 3, m' is 3 and n'' is 0.

在又另一實施例中，n'為4，m'為0且n''為0。在又另一實施例中，n'為4，m'為1且n''為0。在又另一實施例中，n'為4，m'為2且n''為0。在又另一實施例中，n'為4，m'為3且n''為0。In yet another embodiment, n' is 4, m' is 0 and n'' is 0. In yet another embodiment, n' is 4, m' is 1 and n'' is 0. In yet another embodiment, n' is 4, m' is 2 and n'' is 0. In yet another embodiment, n' is 4, m' is 3 and n'' is 0.

在又另一實施例中，n'為5，m'為0且n''為0。在又另一實施例中，n'為5，m'為1且n''為0。在又另一實施例中，n'為5，m'為2且n''為0。在又另一實施例中，n'為5，m'為3且n''為0。In yet another embodiment, n' is 5, m' is 0 and n'' is 0. In yet another embodiment, n' is 5, m' is 1 and n'' is 0. In yet another embodiment, n' is 5, m' is 2 and n'' is 0. In yet another embodiment, n' is 5, m' is 3 and n'' is 0.

在一特定實施例中，n'為1，m'為0且n''為1。在一特定實施例中，n'為1，m'為1且n''為1。在一特定實施例中，n'為1，m'為2且n''為1。在一特定實施例中，n'為1，m'為3且n''為1。In a specific embodiment, n' is 1, m' is 0 and n'' is 1. In a specific embodiment, n' is 1, m' is 1 and n'' is 1. In a specific embodiment, n' is 1, m' is 2 and n'' is 1. In a specific embodiment, n' is 1, m' is 3 and n'' is 1.

在另一實施例中，n'為2，m'為0且n''為1。在另一實施例中，n'為2，m'為1且n''為1。在另一實施例中，n'為2，m'為2且n''為1。在另一實施例中，n'為2，m'為3且n''為1。In another embodiment, n' is 2, m' is 0 and n'' is 1. In another embodiment, n' is 2, m' is 1 and n'' is 1. In another embodiment, n' is 2, m' is 2 and n'' is 1. In another embodiment, n' is 2, m' is 3 and n'' is 1.

在又一實施例中，n'為3，m'為0且n''為1。在又一實施例中，n'為3，m'為1且n''為1。在又一實施例中，n'為3，m'為2且n''為1。在又一實施例中，n'為3，m'為3且n''為1。In yet another embodiment, n' is 3, m' is 0 and n'' is 1. In yet another embodiment, n' is 3, m' is 1 and n'' is 1. In yet another embodiment, n' is 3, m' is 2 and n'' is 1. In yet another embodiment, n' is 3, m' is 3 and n'' is 1.

在又另一實施例中，n'為4，m'為0且n''為1。在又另一實施例中，n'為4，m'為1且n''為1。在又另一實施例中，n'為4，m'為2且n''為1。在又另一實施例中，n'為4，m'為3且n''為1。In yet another embodiment, n' is 4, m' is 0 and n" is 1. In yet another embodiment, n' is 4, m' is 1 and n" is 1. In yet another embodiment, n' is 4, m' is 2 and n" is 1. In yet another embodiment, n' is 4, m' is 3 and n" is 1.

在又另一實施例中，n'為5，m'為0且n''為1。在又另一實施例中，n'為5，m'為1且n''為1。在又另一實施例中，n'為5，m'為2且n''為1。在又另一實施例中，n'為5，m'為3且n''為1。In yet another embodiment, n' is 5, m' is 0 and n'' is 1. In yet another embodiment, n' is 5, m' is 1 and n'' is 1. In yet another embodiment, n' is 5, m' is 2 and n'' is 1. In yet another embodiment, n' is 5, m' is 3 and n'' is 1.

在一特定實施例中，n'為1，m'為0且n''為2。在一特定實施例中，n'為1，m'為1且n''為2。在一特定實施例中，n'為1，m'為2且n''為2。在一特定實施例中，n'為1，m'為3且n''為2。In a specific embodiment, n' is 1, m' is 0 and n'' is 2. In a specific embodiment, n' is 1, m' is 1 and n'' is 2. In a specific embodiment, n' is 1, m' is 2 and n'' is 2. In a specific embodiment, n' is 1, m' is 3 and n'' is 2.

在另一實施例中，n'為2，m'為0且n''為2。在另一實施例中，n'為2，m'為1且n''為2。在另一實施例中，n'為2，m'為2且n''為2。在另一實施例中，n'為2，m'為3且n''為2。In another embodiment, n' is 2, m' is 0 and n'' is 2. In another embodiment, n' is 2, m' is 1 and n'' is 2. In another embodiment, n' is 2, m' is 2 and n'' is 2. In another embodiment, n' is 2, m' is 3 and n'' is 2.

在又一實施例中，n'為3，m'為0且n''為2。在又一實施例中，n'為3，m'為1且n''為2。在又一實施例中，n'為3，m'為2且n''為2。在又一實施例中，n'為3，m'為3且n''為2。In yet another embodiment, n' is 3, m' is 0 and n'' is 2. In yet another embodiment, n' is 3, m' is 1 and n'' is 2. In yet another embodiment, n' is 3, m' is 2 and n'' is 2. In yet another embodiment, n' is 3, m' is 3 and n'' is 2.

在又另一實施例中，n'為4，m'為0且n''為2。在又另一實施例中，n'為4，m'為1且n''為2。在又另一實施例中，n'為4，m'為2且n''為2。在又另一實施例中，n'為4，m'為3且n''為2。In yet another embodiment, n' is 4, m' is 0 and n" is 2. In yet another embodiment, n' is 4, m' is 1 and n" is 2. In yet another embodiment, n' is 4, m' is 2 and n" is 2. In yet another embodiment, n' is 4, m' is 3 and n" is 2.

在又另一實施例中，n'為5，m'為0且n''為2。在又另一實施例中，n'為5，m'為1且n''為2。在又另一實施例中，n'為5，m'為2且n''為2。在又另一實施例中，n'為5，m'為3且n''為2。In yet another embodiment, n' is 5, m' is 0 and n'' is 2. In yet another embodiment, n' is 5, m' is 1 and n'' is 2. In yet another embodiment, n' is 5, m' is 2 and n'' is 2. In yet another embodiment, n' is 5, m' is 3 and n'' is 2.

在一特定實施例中，n'為1，m'為0且n''為3。在一特定實施例中，n'為1，m'為1且n''為3。在一特定實施例中，n'為1，m'為2且n''為3。在一特定實施例中，n'為1，m'為3且n''為3。In a specific embodiment, n' is 1, m' is 0 and n'' is 3. In a specific embodiment, n' is 1, m' is 1 and n'' is 3. In a specific embodiment, n' is 1, m' is 2 and n'' is 3. In a specific embodiment, n' is 1, m' is 3 and n'' is 3.

在另一實施例中，n'為2，m'為0且n''為3。在另一實施例中，n'為2，m'為1且n''為3。在另一實施例中，n'為2，m'為2且n''為3。在另一實施例中，n'為2，m'為3且n''為3。In another embodiment, n' is 2, m' is 0 and n'' is 3. In another embodiment, n' is 2, m' is 1 and n'' is 3. In another embodiment, n' is 2, m' is 2 and n'' is 3. In another embodiment, n' is 2, m' is 3 and n'' is 3.

在又一實施例中，n'為3，m'為0且n''為3。在又一實施例中，n'為3，m'為1且n''為3。在又一實施例中，n'為3，m'為2且n''為3。在又一實施例中，n'為3，m'為3且n''為3。In yet another embodiment, n' is 3, m' is 0 and n'' is 3. In yet another embodiment, n' is 3, m' is 1 and n'' is 3. In yet another embodiment, n' is 3, m' is 2 and n'' is 3. In yet another embodiment, n' is 3, m' is 3 and n'' is 3.

在又另一實施例中，n'為4，m'為0且n''為3。在又另一實施例中，n'為4，m'為1且n''為3。在又另一實施例中，n'為4，m'為2且n''為3。在又另一實施例中，n'為4，m'為3且n''為3。In yet another embodiment, n' is 4, m' is 0 and n'' is 3. In yet another embodiment, n' is 4, m' is 1 and n'' is 3. In yet another embodiment, n' is 4, m' is 2 and n'' is 3. In yet another embodiment, n' is 4, m' is 3 and n'' is 3.

在又另一實施例中，n'為5，m'為0且n''為3。在又另一實施例中，n'為5，m'為1且n''為3。在又另一實施例中，n'為5，m'為2且n''為3。在又另一實施例中，n'為5，m'為3且n''為3。In yet another embodiment, n' is 5, m' is 0 and n'' is 3. In yet another embodiment, n' is 5, m' is 1 and n'' is 3. In yet another embodiment, n' is 5, m' is 2 and n'' is 3. In yet another embodiment, n' is 5, m' is 3 and n'' is 3.

在一特定實施例中，n'為1，m'為0且n''為4。在一特定實施例中，n'為1，m'為1且n''為4。在一特定實施例中，n'為1，m'為2且n''為4。在一特定實施例中，n'為1，m'為3且n''為4。In a specific embodiment, n' is 1, m' is 0 and n'' is 4. In a specific embodiment, n' is 1, m' is 1 and n'' is 4. In a specific embodiment, n' is 1, m' is 2 and n'' is 4. In a specific embodiment, n' is 1, m' is 3 and n'' is 4.

在另一實施例中，n'為2，m'為0且n''為4。在另一實施例中，n'為2，m'為1且n''為4。在另一實施例中，n'為2，m'為2且n''為4。在另一實施例中，n'為2，m'為3且n''為4。In another embodiment, n' is 2, m' is 0 and n'' is 4. In another embodiment, n' is 2, m' is 1 and n'' is 4. In another embodiment, n' is 2, m' is 2 and n'' is 4. In another embodiment, n' is 2, m' is 3 and n'' is 4.

在又一實施例中，n'為3，m'為0且n''為4。在又一實施例中，n'為3，m'為1且n''為4。在又一實施例中，n'為3，m'為2且n''為4。在又一實施例中，n'為3，m'為3且n''為4。In yet another embodiment, n' is 3, m' is 0 and n'' is 4. In yet another embodiment, n' is 3, m' is 1 and n'' is 4. In yet another embodiment, n' is 3, m' is 2 and n'' is 4. In yet another embodiment, n' is 3, m' is 3 and n'' is 4.

在又另一實施例中，n'為4，m'為0且n''為4。在又另一實施例中，n'為4，m'為1且n''為4。在又另一實施例中，n'為4，m'為2且n''為4。在又另一實施例中，n'為4，m'為3且n''為4。In yet another embodiment, n' is 4, m' is 0 and n'' is 4. In yet another embodiment, n' is 4, m' is 1 and n'' is 4. In yet another embodiment, n' is 4, m' is 2 and n'' is 4. In yet another embodiment, n' is 4, m' is 3 and n'' is 4.

在又另一實施例中，n'為5，m'為0且n''為4。在又另一實施例中，n'為5，m'為1且n''為4。在又另一實施例中，n'為5，m'為2且n''為4。在又另一實施例中，n'為5，m'為3且n''為4。In yet another embodiment, n' is 5, m' is 0 and n'' is 4. In yet another embodiment, n' is 5, m' is 1 and n'' is 4. In yet another embodiment, n' is 5, m' is 2 and n'' is 4. In yet another embodiment, n' is 5, m' is 3 and n'' is 4.

在一特定實施例中，n'為1，m'為0且n''為5。在一特定實施例中，n'為1，m'為1且n''為5。在一特定實施例中，n'為1，m'為2且n''為5。在一特定實施例中，n'為1，m'為3且n''為5。In a specific embodiment, n' is 1, m' is 0 and n'' is 5. In a specific embodiment, n' is 1, m' is 1 and n'' is 5. In a specific embodiment, n' is 1, m' is 2 and n'' is 5. In a specific embodiment, n' is 1, m' is 3 and n'' is 5.

在另一實施例中，n'為2，m'為0且n''為5。在另一實施例中，n'為2，m'為1且n''為5。在另一實施例中，n'為2，m'為2且n''為5。在另一實施例中，n'為2，m'為3且n''為5。In another embodiment, n' is 2, m' is 0 and n'' is 5. In another embodiment, n' is 2, m' is 1 and n'' is 5. In another embodiment, n' is 2, m' is 2 and n'' is 5. In another embodiment, n' is 2, m' is 3 and n'' is 5.

在又一實施例中，n'為3，m'為0且n''為5。在又一實施例中，n'為3，m'為1且n''為5。在又一實施例中，n'為3，m'為2且n''為5。在又一實施例中，n'為3，m'為3且n''為5。In yet another embodiment, n' is 3, m' is 0 and n'' is 5. In yet another embodiment, n' is 3, m' is 1 and n'' is 5. In yet another embodiment, n' is 3, m' is 2 and n'' is 5. In yet another embodiment, n' is 3, m' is 3 and n'' is 5.

在又另一實施例中，n'為4，m'為0且n''為5。在又另一實施例中，n'為4，m'為1且n''為5。在又另一實施例中，n'為4，m'為2且n''為5。在又另一實施例中，n'為4，m'為3且n''為5。In yet another embodiment, n' is 4, m' is 0 and n'' is 5. In yet another embodiment, n' is 4, m' is 1 and n'' is 5. In yet another embodiment, n' is 4, m' is 2 and n'' is 5. In yet another embodiment, n' is 4, m' is 3 and n'' is 5.

在又另一實施例中，n'為5，m'為0且n''為5。在又另一實施例中，n'為5，m'為1且n''為5。在又另一實施例中，n'為5，m'為2且n''為5。在又另一實施例中，n'為5，m'為3且n''為5。In yet another embodiment, n' is 5, m' is 0 and n'' is 5. In yet another embodiment, n' is 5, m' is 1 and n'' is 5. In yet another embodiment, n' is 5, m' is 2 and n'' is 5. In yet another embodiment, n' is 5, m' is 3 and n'' is 5.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(IV)，其中X為NHCO(CH 2CH 2O) mCH 2CH 2，其中m係選自1至4，且其中X'為CH 2O(CH 2) n ''CH 2C=O，其中n''係選自0至10。在一實施例中，m係選自1至3且n''係選自0至10。在一實施例中，m係選自1至3且n''係選自0至5。在一實施例中，m係選自1至2且n''係選自0至3。在一實施例中，m係選自1至2且n''係選自0至2。在一特定實施例中，m為1且n''為0。在另一實施例中，m為2且n''為0。在又一實施例中，m為3且n''為0。在又另一實施例中，m為4且n''為0。在一特定實施例中，m為1且n''為1。在另一實施例中，m為2且n''為1。在又一實施例中，m為3且n''為1。在又另一實施例中，m為4且n''為1。在一特定實施例中，m為1且n''為2。在另一實施例中，m為2且n''為2。在又一實施例中，m為3且n''為2。在又另一實施例中，m為4且n''為2。在一特定實施例中，m為1且n''為3。在另一實施例中，m為2且n''為3。在又一實施例中，m為3且n''為3。在又另一實施例中，m為4且n''為3。在一特定實施例中，m為1且n''為4。在另一實施例中，m為2且n''為4。在又一實施例中，m為3且n''為4。在又另一實施例中，m為4且n''為4。在一特定實施例中，m為1且n''為5。在另一實施例中，m為2且n''為5。在又一實施例中，m為3且n''為5。在又另一實施例中，m為4且n''為5。在一特定實施例中，m為1且n''為6。在另一實施例中，m為2且n''為6。在又一實施例中，m為3且n''為6。在又另一實施例中，m為4且n''為6。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (IV), wherein X is NHCO(CH 2 CH 2 O) m CH 2 CH 2 , wherein m is selected from 1 to 4, and wherein X' is CH 2 O(CH 2 ) n '' CH 2 C=O, wherein n '' is selected from 0 to 10. In one embodiment, m is selected from 1 to 3 and n '' is selected from 0 to 10. In one embodiment, m is selected from 1 to 3 and n '' is selected from 0 to 5. In one embodiment, m is selected from 1 to 2 and n '' is selected from 0 to 3. In one embodiment, m is selected from 1 to 2 and n '' is selected from 0 to 2. In a specific embodiment, m is 1 and n '' is 0. In another embodiment, m is 2 and n'' is 0. In yet another embodiment, m is 3 and n'' is 0. In yet another embodiment, m is 4 and n'' is 0. In a specific embodiment, m is 1 and n'' is 1. In another embodiment, m is 2 and n'' is 1. In yet another embodiment, m is 3 and n'' is 1. In yet another embodiment, m is 4 and n'' is 1. In a specific embodiment, m is 1 and n'' is 2. In another embodiment, m is 2 and n'' is 2. In yet another embodiment, m is 3 and n'' is 2. In yet another embodiment, m is 4 and n'' is 2. In a specific embodiment, m is 1 and n'' is 3. In another embodiment, m is 2 and n'' is 3. In yet another embodiment, m is 3 and n'' is 3. In yet another embodiment, m is 4 and n'' is 3. In a specific embodiment, m is 1 and n'' is 4. In another embodiment, m is 2 and n'' is 4. In yet another embodiment, m is 3 and n'' is 4. In yet another embodiment, m is 4 and n'' is 4. In a specific embodiment, m is 1 and n'' is 5. In another embodiment, m is 2 and n'' is 5. In yet another embodiment, m is 3 and n'' is 5. In yet another embodiment, m is 4 and n'' is 5. In a specific embodiment, m is 1 and n'' is 6. In another embodiment, m is 2 and n'' is 6. In yet another embodiment, m is 3 and n'' is 6. In yet another embodiment, m is 4 and n″ is 6.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(IV)，其中X為NHCO(CH 2CH 2O) mCH 2CH 2，其中m係選自1至4，且其中X'為CH 2O(CH 2CH 2O) m '(CH 2) n ''CH 2C=O，其中n''係選自0至10且m'係選自0至4。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (IV), wherein X is NHCO(CH 2 CH 2 O) m CH 2 CH 2 , wherein m is selected from 1 to 4, and wherein X' is CH 2 O(CH 2 CH 2 O) m ' (CH 2 ) n '' CH 2 C=O, wherein n'' is selected from 0 to 10 and m' is selected from 0 to 4.

在一實施例中，m係選自1至3，m'係選自0至4且n''係選自0至10。在一實施例中，m係選自1至2，m'係選自0至4且n''係選自0至5。在一實施例中，m係選自1至2，m'係選自0至2且n''係選自0至3。在一實施例中，m係選自1至2，m'係選自0至2且n''係選自0至1。In one embodiment, m is selected from 1 to 3, m' is selected from 0 to 4 and n'' is selected from 0 to 10. In one embodiment, m is selected from 1 to 2, m' is selected from 0 to 4 and n'' is selected from 0 to 5. In one embodiment, m is selected from 1 to 2, m' is selected from 0 to 2 and n'' is selected from 0 to 3. In one embodiment, m is selected from 1 to 2, m' is selected from 0 to 2 and n'' is selected from 0 to 1.

在一特定實施例中，m為1，m'為0且n''為0。在另一實施例中，m為1，m'為1且n''為0。在另一實施例中，m為1，m'為2且n''為0。在另一實施例中，m為1，m'為3且n''為0。In a particular embodiment, m is 1, m' is 0 and n'' is 0. In another embodiment, m is 1, m' is 1 and n'' is 0. In another embodiment, m is 1, m' is 2 and n'' is 0. In another embodiment, m is 1, m' is 3 and n'' is 0.

在另一實施例中，m為2，m'為0且n''為0。在另一實施例中，m為2，m'為1且n''為0。在另一實施例中，m為2，m'為2且n''為0。在另一實施例中，m為2，m'為3且n''為0。In another embodiment, m is 2, m' is 0 and n'' is 0. In another embodiment, m is 2, m' is 1 and n'' is 0. In another embodiment, m is 2, m' is 2 and n'' is 0. In another embodiment, m is 2, m' is 3 and n'' is 0.

在又一實施例中，m為3，m'為0且n''為0。在又一實施例中，m為3，m'為1且n''為0。在又一實施例中，m為3，m'為2且n''為0。在又一實施例中，m為3，m'為3且n''為0。In yet another embodiment, m is 3, m' is 0 and n'' is 0. In yet another embodiment, m is 3, m' is 1 and n'' is 0. In yet another embodiment, m is 3, m' is 2 and n'' is 0. In yet another embodiment, m is 3, m' is 3 and n'' is 0.

在又另一實施例中，m為4，m'為0且n''為0。在又另一實施例中，m為4，m'為1且n''為0。在又另一實施例中，m為4，m'為2且n''為0。在又另一實施例中，m為4，m'為3且n''為0。In yet another embodiment, m is 4, m' is 0 and n" is 0. In yet another embodiment, m is 4, m' is 1 and n" is 0. In yet another embodiment, m is 4, m' is 2 and n" is 0. In yet another embodiment, m is 4, m' is 3 and n" is 0.

在一特定實施例中，m為1，m'為0且n''為1。在一特定實施例中，m為1，m'為1且n''為1。在一特定實施例中，m為1，m'為2且n''為1。在一特定實施例中，m為1，m'為3且n''為1。In a specific embodiment, m is 1, m' is 0 and n'' is 1. In a specific embodiment, m is 1, m' is 1 and n'' is 1. In a specific embodiment, m is 1, m' is 2 and n'' is 1. In a specific embodiment, m is 1, m' is 3 and n'' is 1.

在另一實施例中，m為2，m'為0且n''為1。在另一實施例中，m為2，m'為1且n''為1。在另一實施例中，m為2，m'為2且n''為1。在另一實施例中，m為2，m'為3且n''為1。In another embodiment, m is 2, m' is 0 and n'' is 1. In another embodiment, m is 2, m' is 1 and n'' is 1. In another embodiment, m is 2, m' is 2 and n'' is 1. In another embodiment, m is 2, m' is 3 and n'' is 1.

在又一實施例中，m為3，m'為0且n''為1。在又一實施例中，m為3，m'為1且n''為1。在又一實施例中，m為3，m'為2且n''為1。在又一實施例中，m為3，m'為3且n''為1。In yet another embodiment, m is 3, m' is 0 and n'' is 1. In yet another embodiment, m is 3, m' is 1 and n'' is 1. In yet another embodiment, m is 3, m' is 2 and n'' is 1. In yet another embodiment, m is 3, m' is 3 and n'' is 1.

在又另一實施例中，m為4，m'為0且n''為1。在又另一實施例中，m為4，m'為1且n''為1。在又另一實施例中，m為4，m'為2且n''為1。在又另一實施例中，m為4，m'為3且n''為1。In yet another embodiment, m is 4, m' is 0 and n'' is 1. In yet another embodiment, m is 4, m' is 1 and n'' is 1. In yet another embodiment, m is 4, m' is 2 and n'' is 1. In yet another embodiment, m is 4, m' is 3 and n'' is 1.

在一特定實施例中，m為1，m'為0且n''為2。在一特定實施例中，m為1，m'為1且n''為2。在一特定實施例中，m為1，m'為2且n''為2。在一特定實施例中，m為1，m'為3且n''為2。In a specific embodiment, m is 1, m' is 0 and n'' is 2. In a specific embodiment, m is 1, m' is 1 and n'' is 2. In a specific embodiment, m is 1, m' is 2 and n'' is 2. In a specific embodiment, m is 1, m' is 3 and n'' is 2.

在另一實施例中，m為2，m'為0且n''為2。在另一實施例中，m為2，m'為1且n''為2。在另一實施例中，m為2，m'為2且n''為2。在另一實施例中，m為2，m'為3且n''為2。In another embodiment, m is 2, m' is 0 and n'' is 2. In another embodiment, m is 2, m' is 1 and n'' is 2. In another embodiment, m is 2, m' is 2 and n'' is 2. In another embodiment, m is 2, m' is 3 and n'' is 2.

在又一實施例中，m為3，m'為0且n''為2。在又一實施例中，m為3，m'為1且n''為2。在又一實施例中，m為3，m'為2且n''為2。在又一實施例中，m為3，m'為3且n''為2。In yet another embodiment, m is 3, m' is 0 and n'' is 2. In yet another embodiment, m is 3, m' is 1 and n'' is 2. In yet another embodiment, m is 3, m' is 2 and n'' is 2. In yet another embodiment, m is 3, m' is 3 and n'' is 2.

在又另一實施例中，m為4，m'為0且n''為2。在又另一實施例中，m為4，m'為1且n''為2。在又另一實施例中，m為4，m'為2且n''為2。在又另一實施例中，m為4，m'為3且n''為2。In yet another embodiment, m is 4, m' is 0 and n'' is 2. In yet another embodiment, m is 4, m' is 1 and n'' is 2. In yet another embodiment, m is 4, m' is 2 and n'' is 2. In yet another embodiment, m is 4, m' is 3 and n'' is 2.

在一特定實施例中，m為1，m'為0且n''為3。在一特定實施例中，m為1，m'為1且n''為3。在一特定實施例中，m為1，m'為2且n''為3。在一特定實施例中，m為1，m'為3且n''為3。In a specific embodiment, m is 1, m' is 0 and n'' is 3. In a specific embodiment, m is 1, m' is 1 and n'' is 3. In a specific embodiment, m is 1, m' is 2 and n'' is 3. In a specific embodiment, m is 1, m' is 3 and n'' is 3.

在另一實施例中，m為2，m'為0且n''為3。在另一實施例中，m為2，m'為1且n''為3。在另一實施例中，m為2，m'為2且n''為3。在另一實施例中，m為2，m'為3且n''為3。In another embodiment, m is 2, m' is 0 and n'' is 3. In another embodiment, m is 2, m' is 1 and n'' is 3. In another embodiment, m is 2, m' is 2 and n'' is 3. In another embodiment, m is 2, m' is 3 and n'' is 3.

在又一實施例中，m為3，m'為0且n''為3。在又一實施例中，m為3，m'為1且n''為3。在又一實施例中，m為3，m'為2且n''為3。在又一實施例中，m為3，m'為3且n''為3。In yet another embodiment, m is 3, m' is 0 and n'' is 3. In yet another embodiment, m is 3, m' is 1 and n'' is 3. In yet another embodiment, m is 3, m' is 2 and n'' is 3. In yet another embodiment, m is 3, m' is 3 and n'' is 3.

在又另一實施例中，m為4，m'為0且n''為3。在又另一實施例中，m為4，m'為1且n''為3。在又另一實施例中，m為4，m'為2且n''為3。在又另一實施例中，m為4，m'為3且n''為3。In yet another embodiment, m is 4, m' is 0 and n'' is 3. In yet another embodiment, m is 4, m' is 1 and n'' is 3. In yet another embodiment, m is 4, m' is 2 and n'' is 3. In yet another embodiment, m is 4, m' is 3 and n'' is 3.

在一特定實施例中，m為1，m'為0且n''為4。在一特定實施例中，m為1，m'為1且n''為4。在一特定實施例中，m為1，m'為2且n''為4。在一特定實施例中，m為1，m'為3且n''為4。In a specific embodiment, m is 1, m' is 0 and n'' is 4. In a specific embodiment, m is 1, m' is 1 and n'' is 4. In a specific embodiment, m is 1, m' is 2 and n'' is 4. In a specific embodiment, m is 1, m' is 3 and n'' is 4.

在另一實施例中，m為2，m'為0且n''為4。在另一實施例中，m為2，m'為1且n''為4。在另一實施例中，m為2，m'為2且n''為4。在另一實施例中，m為2，m'為3且n''為4。In another embodiment, m is 2, m' is 0 and n'' is 4. In another embodiment, m is 2, m' is 1 and n'' is 4. In another embodiment, m is 2, m' is 2 and n'' is 4. In another embodiment, m is 2, m' is 3 and n'' is 4.

在又一實施例中，m為3，m'為0且n''為4。在又一實施例中，m為3，m'為1且n''為4。在又一實施例中，m為3，m'為2且n''為4。在又一實施例中，m為3，m'為3且n''為4。In yet another embodiment, m is 3, m' is 0 and n'' is 4. In yet another embodiment, m is 3, m' is 1 and n'' is 4. In yet another embodiment, m is 3, m' is 2 and n'' is 4. In yet another embodiment, m is 3, m' is 3 and n'' is 4.

在又另一實施例中，m為4，m'為0且n''為4。在又另一實施例中，m為4，m'為1且n''為4。在又另一實施例中，m為4，m'為2且n''為4。在又另一實施例中，m為4，m'為3且n''為4。In yet another embodiment, m is 4, m' is 0 and n'' is 4. In yet another embodiment, m is 4, m' is 1 and n'' is 4. In yet another embodiment, m is 4, m' is 2 and n'' is 4. In yet another embodiment, m is 4, m' is 3 and n'' is 4.

在一特定實施例中，m為1，m'為0且n''為5。在一特定實施例中，m為1，m'為1且n''為5。在一特定實施例中，m為1，m'為2且n''為5。在一特定實施例中，m為1，m'為3且n''為5。In a specific embodiment, m is 1, m' is 0 and n'' is 5. In a specific embodiment, m is 1, m' is 1 and n'' is 5. In a specific embodiment, m is 1, m' is 2 and n'' is 5. In a specific embodiment, m is 1, m' is 3 and n'' is 5.

在另一實施例中，m為2，m'為0且n''為5。在另一實施例中，m為2，m'為1且n''為5。在另一實施例中，m為2，m'為2且n''為5。在另一實施例中，m為2，m'為3且n''為5。In another embodiment, m is 2, m' is 0 and n'' is 5. In another embodiment, m is 2, m' is 1 and n'' is 5. In another embodiment, m is 2, m' is 2 and n'' is 5. In another embodiment, m is 2, m' is 3 and n'' is 5.

在又一實施例中，m為3，m'為0且n''為5。在又一實施例中，m為3，m'為1且n''為5。在又一實施例中，m為3，m'為2且n''為5。在又一實施例中，m為3，m'為3且n''為5。In yet another embodiment, m is 3, m' is 0 and n'' is 5. In yet another embodiment, m is 3, m' is 1 and n'' is 5. In yet another embodiment, m is 3, m' is 2 and n'' is 5. In yet another embodiment, m is 3, m' is 3 and n'' is 5.

在又另一實施例中，m為4，m'為0且n''為5。在又另一實施例中，m為4，m'為1且n''為5。在又另一實施例中，m為4，m'為2且n''為5。在又另一實施例中，m為4，m'為3且n''為5。In yet another embodiment, m is 4, m' is 0 and n'' is 5. In yet another embodiment, m is 4, m' is 1 and n'' is 5. In yet another embodiment, m is 4, m' is 2 and n'' is 5. In yet another embodiment, m is 4, m' is 3 and n'' is 5.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(IV)，其中X為OCH 2(CH 2) n '，其中n'係選自1至10，且其中X'為CH 2O(CH 2) n ''CH 2C=O，其中n''係選自0至10。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (IV), wherein X is OCH2 ( CH2 ) n ' , wherein n' is selected from 1 to 10, and wherein X' is CH2O ( CH2 )n''CH2C = O , wherein n'' is selected from 0 to 10.

在一實施例中，n'係選自1至5且n''係選自0至10。在一實施例中，n'係選自1至5且n''係選自0至5。在一實施例中，n'係選自1至3且n''係選自0至3。在一實施例中，n'係選自1至2且n''係選自0至2。在一特定實施例中，n'為1且n''為0。在另一實施例中，n'為2且n''為0。在又一實施例中，n'為3且n''為0。在又另一實施例中，n'為4且n''為0。在又另一實施例中，n'為5且n''為0。在又另一實施例中，n'為6且n''為0。在一特定實施例中，n'為1且n''為1。在另一實施例中，n'為2且n''為1。在又一實施例中，n'為3且n''為1。在又另一實施例中，n'為4且n''為1。在又另一實施例中，n'為5且n''為1。在又另一實施例中，n'為6且n''為1。在一特定實施例中，n'為1且n''為2。在另一實施例中，n'為2且n''為2。在又一實施例中，n'為3且n''為2。在又另一實施例中，n'為4且n''為2。在又另一實施例中，n'為5且n''為2。在又另一實施例中，n'為6且n''為2。在一特定實施例中，n'為1且n''為3。在另一實施例中，n'為2且n''為3。在又一實施例中，n'為3且n''為3。在又另一實施例中，n'為4且n''為3。在又另一實施例中，n'為5且n''為3。在又另一實施例中，n'為6且n''為3。在一特定實施例中，n'為1且n''為4。在另一實施例中，n'為2且n''為4。在又一實施例中，n'為3且n''為4。在又另一實施例中，n'為4且n''為4。在又另一實施例中，n'為5且n''為4。在又另一實施例中，n'為6且n''為4。在一特定實施例中，n'為1且n''為5。在另一實施例中，n'為2且n''為5。在又一實施例中，n'為3且n''為5。在又另一實施例中，n'為4且n''為5。在又另一實施例中，n'為5且n''為5。在又另一實施例中，n'為6且n''為5。在一特定實施例中，n'為1且n''為6。在另一實施例中，n'為2且n''為6。在又一實施例中，n'為3且n''為6。在又另一實施例中，n'為4且n''為6。在又另一實施例中，n'為5且n''為6。在又另一實施例中，n'為6且n''為6。In one embodiment, n' is selected from 1 to 5 and n'' is selected from 0 to 10. In one embodiment, n' is selected from 1 to 5 and n'' is selected from 0 to 5. In one embodiment, n' is selected from 1 to 3 and n'' is selected from 0 to 3. In one embodiment, n' is selected from 1 to 2 and n'' is selected from 0 to 2. In a specific embodiment, n' is 1 and n'' is 0. In another embodiment, n' is 2 and n'' is 0. In yet another embodiment, n' is 3 and n'' is 0. In yet another embodiment, n' is 4 and n'' is 0. In yet another embodiment, n' is 5 and n'' is 0. In yet another embodiment, n' is 6 and n'' is 0. In a specific embodiment, n' is 1 and n'' is 1. In another embodiment, n' is 2 and n'' is 1. In yet another embodiment, n' is 3 and n'' is 1. In yet another embodiment, n' is 4 and n'' is 1. In yet another embodiment, n' is 5 and n'' is 1. In yet another embodiment, n' is 6 and n'' is 1. In a specific embodiment, n' is 1 and n'' is 2. In another embodiment, n' is 2 and n'' is 2. In yet another embodiment, n' is 3 and n'' is 2. In yet another embodiment, n' is 4 and n'' is 2. In yet another embodiment, n' is 5 and n'' is 2. In yet another embodiment, n' is 6 and n'' is 2. In a specific embodiment, n' is 1 and n'' is 3. In another embodiment, n' is 2 and n'' is 3. In yet another embodiment, n' is 3 and n'' is 3. In yet another embodiment, n' is 4 and n'' is 3. In yet another embodiment, n' is 5 and n'' is 3. In yet another embodiment, n' is 6 and n'' is 3. In a specific embodiment, n' is 1 and n'' is 4. In another embodiment, n' is 2 and n'' is 4. In yet another embodiment, n' is 3 and n'' is 4. In yet another embodiment, n' is 4 and n'' is 4. In yet another embodiment, n' is 5 and n'' is 4. In yet another embodiment, n' is 6 and n'' is 4. In a specific embodiment, n' is 1 and n'' is 5. In another embodiment, n' is 2 and n'' is 5. In yet another embodiment, n' is 3 and n'' is 5. In yet another embodiment, n' is 4 and n" is 5. In yet another embodiment, n' is 5 and n" is 5. In yet another embodiment, n' is 6 and n" is 5. In a particular embodiment, n' is 1 and n" is 6. In another embodiment, n' is 2 and n" is 6. In yet another embodiment, n' is 3 and n" is 6. In yet another embodiment, n' is 4 and n" is 6. In yet another embodiment, n' is 5 and n" is 6. In yet another embodiment, n' is 6 and n" is 6.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(IV)，其中X為OCH 2(CH 2) n '，其中n'係選自1至10，且其中X'為CH 2O(CH 2CH 2O) m '(CH 2) n ''CH 2C=O，其中n''係選自0至10且m'係選自0至4。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (IV), wherein X is OCH2 ( CH2 ) n ' , wherein n' is selected from 1 to 10, and wherein X ' is CH2O ( CH2CH2O ) m ' ( CH2 ) n''CH2C = O , wherein n'' is selected from 0 to 10 and m' is selected from 0 to 4.

在一實施例中，n'係選自1至5，m'係選自0至4且n''係選自0至10。在一實施例中，n'係選自1至5，m'係選自0至4且n''係選自0至5。在一實施例中，n'係選自1至3，m'係選自0至2且n''係選自0至3。在一實施例中，n'係選自1至2，m'係選自0至2且n''係選自0至1。In one embodiment, n' is selected from 1 to 5, m' is selected from 0 to 4 and n'' is selected from 0 to 10. In one embodiment, n' is selected from 1 to 5, m' is selected from 0 to 4 and n'' is selected from 0 to 5. In one embodiment, n' is selected from 1 to 3, m' is selected from 0 to 2 and n'' is selected from 0 to 3. In one embodiment, n' is selected from 1 to 2, m' is selected from 0 to 2 and n'' is selected from 0 to 1.

在一特定實施例中，n'為1，m'為0且n''為0。在另一實施例中，n'為1，m'為1且n''為0。在另一實施例中，n'為1，m'為2且n''為0。在另一實施例中，n'為1，m'為3且n''為0。In a particular embodiment, n' is 1, m' is 0 and n'' is 0. In another embodiment, n' is 1, m' is 1 and n'' is 0. In another embodiment, n' is 1, m' is 2 and n'' is 0. In another embodiment, n' is 1, m' is 3 and n'' is 0.

在另一實施例中，n'為2，m'為0且n''為0。在另一實施例中，n'為2，m'為1且n''為0。在另一實施例中，n'為2，m'為2且n''為0。在另一實施例中，n'為2，m'為3且n''為0。In another embodiment, n' is 2, m' is 0 and n'' is 0. In another embodiment, n' is 2, m' is 1 and n'' is 0. In another embodiment, n' is 2, m' is 2 and n'' is 0. In another embodiment, n' is 2, m' is 3 and n'' is 0.

在又一實施例中，n'為3，m'為0且n''為0。在又一實施例中，n'為3，m'為1且n''為0。在又一實施例中，n'為3，m'為2且n''為0。在又一實施例中，n'為3，m'為3且n''為0。In yet another embodiment, n' is 3, m' is 0 and n'' is 0. In yet another embodiment, n' is 3, m' is 1 and n'' is 0. In yet another embodiment, n' is 3, m' is 2 and n'' is 0. In yet another embodiment, n' is 3, m' is 3 and n'' is 0.

在又另一實施例中，n'為4，m'為0且n''為0。在又另一實施例中，n'為4，m'為1且n''為0。在又另一實施例中，n'為4，m'為2且n''為0。在又另一實施例中，n'為4，m'為3且n''為0。In yet another embodiment, n' is 4, m' is 0 and n'' is 0. In yet another embodiment, n' is 4, m' is 1 and n'' is 0. In yet another embodiment, n' is 4, m' is 2 and n'' is 0. In yet another embodiment, n' is 4, m' is 3 and n'' is 0.

在又另一實施例中，n'為5，m'為0且n''為0。在又另一實施例中，n'為5，m'為1且n''為0。在又另一實施例中，n'為5，m'為2且n''為0。在又另一實施例中，n'為5，m'為3且n''為0。In yet another embodiment, n' is 5, m' is 0 and n'' is 0. In yet another embodiment, n' is 5, m' is 1 and n'' is 0. In yet another embodiment, n' is 5, m' is 2 and n'' is 0. In yet another embodiment, n' is 5, m' is 3 and n'' is 0.

在一特定實施例中，n'為1，m'為0且n''為1。在一特定實施例中，n'為1，m'為1且n''為1。在一特定實施例中，n'為1，m'為2且n''為1。在一特定實施例中，n'為1，m'為3且n''為1。In a specific embodiment, n' is 1, m' is 0 and n'' is 1. In a specific embodiment, n' is 1, m' is 1 and n'' is 1. In a specific embodiment, n' is 1, m' is 2 and n'' is 1. In a specific embodiment, n' is 1, m' is 3 and n'' is 1.

在另一實施例中，n'為2，m'為0且n''為1。在另一實施例中，n'為2，m'為1且n''為1。在另一實施例中，n'為2，m'為2且n''為1。在另一實施例中，n'為2，m'為3且n''為1。In another embodiment, n' is 2, m' is 0 and n'' is 1. In another embodiment, n' is 2, m' is 1 and n'' is 1. In another embodiment, n' is 2, m' is 2 and n'' is 1. In another embodiment, n' is 2, m' is 3 and n'' is 1.

在又一實施例中，n'為3，m'為0且n''為1。在又一實施例中，n'為3，m'為1且n''為1。在又一實施例中，n'為3，m'為2且n''為1。在又一實施例中，n'為3，m'為3且n''為1。In yet another embodiment, n' is 3, m' is 0 and n'' is 1. In yet another embodiment, n' is 3, m' is 1 and n'' is 1. In yet another embodiment, n' is 3, m' is 2 and n'' is 1. In yet another embodiment, n' is 3, m' is 3 and n'' is 1.

在又另一實施例中，n'為4，m'為0且n''為1。在又另一實施例中，n'為4，m'為1且n''為1。在又另一實施例中，n'為4，m'為2且n''為1。在又另一實施例中，n'為4，m'為3且n''為1。In yet another embodiment, n' is 4, m' is 0 and n" is 1. In yet another embodiment, n' is 4, m' is 1 and n" is 1. In yet another embodiment, n' is 4, m' is 2 and n" is 1. In yet another embodiment, n' is 4, m' is 3 and n" is 1.

在又另一實施例中，n'為5，m'為0且n''為1。在又另一實施例中，n'為5，m'為1且n''為1。在又另一實施例中，n'為5，m'為2且n''為1。在又另一實施例中，n'為5，m'為3且n''為1。In yet another embodiment, n' is 5, m' is 0 and n'' is 1. In yet another embodiment, n' is 5, m' is 1 and n'' is 1. In yet another embodiment, n' is 5, m' is 2 and n'' is 1. In yet another embodiment, n' is 5, m' is 3 and n'' is 1.

在一特定實施例中，n'為1，m'為0且n''為2。在一特定實施例中，n'為1，m'為1且n''為2。在一特定實施例中，n'為1，m'為2且n''為2。在一特定實施例中，n'為1，m'為3且n''為2。In a specific embodiment, n' is 1, m' is 0 and n'' is 2. In a specific embodiment, n' is 1, m' is 1 and n'' is 2. In a specific embodiment, n' is 1, m' is 2 and n'' is 2. In a specific embodiment, n' is 1, m' is 3 and n'' is 2.

在另一實施例中，n'為2，m'為0且n''為2。在另一實施例中，n'為2，m'為1且n''為2。在另一實施例中，n'為2，m'為2且n''為2。在另一實施例中，n'為2，m'為3且n''為2。In another embodiment, n' is 2, m' is 0 and n'' is 2. In another embodiment, n' is 2, m' is 1 and n'' is 2. In another embodiment, n' is 2, m' is 2 and n'' is 2. In another embodiment, n' is 2, m' is 3 and n'' is 2.

在又一實施例中，n'為3，m'為0且n''為2。在又一實施例中，n'為3，m'為1且n''為2。在又一實施例中，n'為3，m'為2且n''為2。在又一實施例中，n'為3，m'為3且n''為2。In yet another embodiment, n' is 3, m' is 0 and n'' is 2. In yet another embodiment, n' is 3, m' is 1 and n'' is 2. In yet another embodiment, n' is 3, m' is 2 and n'' is 2. In yet another embodiment, n' is 3, m' is 3 and n'' is 2.

在又另一實施例中，n'為4，m'為0且n''為2。在又另一實施例中，n'為4，m'為1且n''為2。在又另一實施例中，n'為4，m'為2且n''為2。在又另一實施例中，n'為4，m'為3且n''為2。In yet another embodiment, n' is 4, m' is 0 and n" is 2. In yet another embodiment, n' is 4, m' is 1 and n" is 2. In yet another embodiment, n' is 4, m' is 2 and n" is 2. In yet another embodiment, n' is 4, m' is 3 and n" is 2.

在又另一實施例中，n'為5，m'為0且n''為2。在又另一實施例中，n'為5，m'為1且n''為2。在又另一實施例中，n'為5，m'為2且n''為2。在又另一實施例中，n'為5，m'為3且n''為2。In yet another embodiment, n' is 5, m' is 0 and n'' is 2. In yet another embodiment, n' is 5, m' is 1 and n'' is 2. In yet another embodiment, n' is 5, m' is 2 and n'' is 2. In yet another embodiment, n' is 5, m' is 3 and n'' is 2.

在一特定實施例中，n'為1，m'為0且n''為3。在一特定實施例中，n'為1，m'為1且n''為3。在一特定實施例中，n'為1，m'為2且n''為3。在一特定實施例中，n'為1，m'為3且n''為3。In a specific embodiment, n' is 1, m' is 0 and n'' is 3. In a specific embodiment, n' is 1, m' is 1 and n'' is 3. In a specific embodiment, n' is 1, m' is 2 and n'' is 3. In a specific embodiment, n' is 1, m' is 3 and n'' is 3.

在另一實施例中，n'為2，m'為0且n''為3。在另一實施例中，n'為2，m'為1且n''為3。在另一實施例中，n'為2，m'為2且n''為3。在另一實施例中，n'為2，m'為3且n''為3。In another embodiment, n' is 2, m' is 0 and n'' is 3. In another embodiment, n' is 2, m' is 1 and n'' is 3. In another embodiment, n' is 2, m' is 2 and n'' is 3. In another embodiment, n' is 2, m' is 3 and n'' is 3.

在又一實施例中，n'為3，m'為0且n''為3。在又一實施例中，n'為3，m'為1且n''為3。在又一實施例中，n'為3，m'為2且n''為3。在又一實施例中，n'為3，m'為3且n''為3。In yet another embodiment, n' is 3, m' is 0 and n'' is 3. In yet another embodiment, n' is 3, m' is 1 and n'' is 3. In yet another embodiment, n' is 3, m' is 2 and n'' is 3. In yet another embodiment, n' is 3, m' is 3 and n'' is 3.

在又另一實施例中，n'為4，m'為0且n''為3。在又另一實施例中，n'為4，m'為1且n''為3。在又另一實施例中，n'為4，m'為2且n''為3。在又另一實施例中，n'為4，m'為3且n''為3。In yet another embodiment, n' is 4, m' is 0 and n'' is 3. In yet another embodiment, n' is 4, m' is 1 and n'' is 3. In yet another embodiment, n' is 4, m' is 2 and n'' is 3. In yet another embodiment, n' is 4, m' is 3 and n'' is 3.

在又另一實施例中，n'為5，m'為0且n''為3。在又另一實施例中，n'為5，m'為1且n''為3。在又另一實施例中，n'為5，m'為2且n''為3。在又另一實施例中，n'為5，m'為3且n''為3。In yet another embodiment, n' is 5, m' is 0 and n'' is 3. In yet another embodiment, n' is 5, m' is 1 and n'' is 3. In yet another embodiment, n' is 5, m' is 2 and n'' is 3. In yet another embodiment, n' is 5, m' is 3 and n'' is 3.

在一特定實施例中，n'為1，m'為0且n''為4。在一特定實施例中，n'為1，m'為1且n''為4。在一特定實施例中，n'為1，m'為2且n''為4。在一特定實施例中，n'為1，m'為3且n''為4。In a specific embodiment, n' is 1, m' is 0 and n'' is 4. In a specific embodiment, n' is 1, m' is 1 and n'' is 4. In a specific embodiment, n' is 1, m' is 2 and n'' is 4. In a specific embodiment, n' is 1, m' is 3 and n'' is 4.

在另一實施例中，n'為2，m'為0且n''為4。在另一實施例中，n'為2，m'為1且n''為4。在另一實施例中，n'為2，m'為2且n''為4。在另一實施例中，n'為2，m'為3且n''為4。In another embodiment, n' is 2, m' is 0 and n'' is 4. In another embodiment, n' is 2, m' is 1 and n'' is 4. In another embodiment, n' is 2, m' is 2 and n'' is 4. In another embodiment, n' is 2, m' is 3 and n'' is 4.

在又一實施例中，n'為3，m'為0且n''為4。在又一實施例中，n'為3，m'為1且n''為4。在又一實施例中，n'為3，m'為2且n''為4。在又一實施例中，n'為3，m'為3且n''為4。In yet another embodiment, n' is 3, m' is 0 and n'' is 4. In yet another embodiment, n' is 3, m' is 1 and n'' is 4. In yet another embodiment, n' is 3, m' is 2 and n'' is 4. In yet another embodiment, n' is 3, m' is 3 and n'' is 4.

在又另一實施例中，n'為4，m'為0且n''為4。在又另一實施例中，n'為4，m'為1且n''為4。在又另一實施例中，n'為4，m'為2且n''為4。在又另一實施例中，n'為4，m'為3且n''為4。In yet another embodiment, n' is 4, m' is 0 and n'' is 4. In yet another embodiment, n' is 4, m' is 1 and n'' is 4. In yet another embodiment, n' is 4, m' is 2 and n'' is 4. In yet another embodiment, n' is 4, m' is 3 and n'' is 4.

在又另一實施例中，n'為5，m'為0且n''為4。在又另一實施例中，n'為5，m'為1且n''為4。在又另一實施例中，n'為5，m'為2且n''為4。在又另一實施例中，n'為5，m'為3且n''為4。In yet another embodiment, n' is 5, m' is 0 and n'' is 4. In yet another embodiment, n' is 5, m' is 1 and n'' is 4. In yet another embodiment, n' is 5, m' is 2 and n'' is 4. In yet another embodiment, n' is 5, m' is 3 and n'' is 4.

在一特定實施例中，n'為1，m'為0且n''為5。在一特定實施例中，n'為1，m'為1且n''為5。在一特定實施例中，n'為1，m'為2且n''為5。在一特定實施例中，n'為1，m'為3且n''為5。In a specific embodiment, n' is 1, m' is 0 and n'' is 5. In a specific embodiment, n' is 1, m' is 1 and n'' is 5. In a specific embodiment, n' is 1, m' is 2 and n'' is 5. In a specific embodiment, n' is 1, m' is 3 and n'' is 5.

在另一實施例中，n'為2，m'為0且n''為5。在另一實施例中，n'為2，m'為1且n''為5。在另一實施例中，n'為2，m'為2且n''為5。在另一實施例中，n'為2，m'為3且n''為5。In another embodiment, n' is 2, m' is 0 and n'' is 5. In another embodiment, n' is 2, m' is 1 and n'' is 5. In another embodiment, n' is 2, m' is 2 and n'' is 5. In another embodiment, n' is 2, m' is 3 and n'' is 5.

在又一實施例中，n'為3，m'為0且n''為5。在又一實施例中，n'為3，m'為1且n''為5。在又一實施例中，n'為3，m'為2且n''為5。在又一實施例中，n'為3，m'為3且n''為5。In yet another embodiment, n' is 3, m' is 0 and n'' is 5. In yet another embodiment, n' is 3, m' is 1 and n'' is 5. In yet another embodiment, n' is 3, m' is 2 and n'' is 5. In yet another embodiment, n' is 3, m' is 3 and n'' is 5.

在又另一實施例中，n'為4，m'為0且n''為5。在又另一實施例中，n'為4，m'為1且n''為5。在又另一實施例中，n'為4，m'為2且n''為5。在又另一實施例中，n'為4，m'為3且n''為5。In yet another embodiment, n' is 4, m' is 0 and n'' is 5. In yet another embodiment, n' is 4, m' is 1 and n'' is 5. In yet another embodiment, n' is 4, m' is 2 and n'' is 5. In yet another embodiment, n' is 4, m' is 3 and n'' is 5.

在又另一實施例中，n'為5，m'為0且n''為5。在又另一實施例中，n'為5，m'為1且n''為5。在又另一實施例中，n'為5，m'為2且n''為5。在又另一實施例中，n'為5，m'為3且n''為5。In yet another embodiment, n' is 5, m' is 0 and n'' is 5. In yet another embodiment, n' is 5, m' is 1 and n'' is 5. In yet another embodiment, n' is 5, m' is 2 and n'' is 5. In yet another embodiment, n' is 5, m' is 3 and n'' is 5.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(IV)，其中X為O(CH 2CH 2O) mCH 2CH 2，其中m係選自1至4，且其中X'為CH 2O(CH 2) n ''CH 2C=O，其中n''係選自0至10。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (IV), wherein X is O(CH 2 CH 2 O) m CH 2 CH 2 , wherein m is selected from 1 to 4, and wherein X' is CH 2 O(CH 2 ) n '' CH 2 C=O, wherein n'' is selected from 0 to 10.

在一實施例中，m係選自1至3且n''係選自0至10。在一實施例中，m係選自1至3且n''係選自0至5。在一實施例中，m係選自1至2且n''係選自0至3。在一實施例中，m係選自1至2且n''係選自0至2。在一特定實施例中，m為1且n''為0。在另一實施例中，m為2且n''為0。在又一實施例中，m為3且n''為0。在又另一實施例中，m為4且n''為0。在一特定實施例中，m為1且n''為1。在另一實施例中，m為2且n''為1。在又一實施例中，m為3且n''為1。在又另一實施例中，m為4且n''為1。在一特定實施例中，m為1且n''為2。在另一實施例中，m為2且n''為2。在又一實施例中，m為3且n''為2。在又另一實施例中，m為4且n''為2。在一特定實施例中，m為1且n''為3。在另一實施例中，m為2且n''為3。在又一實施例中，m為3且n''為3。在又另一實施例中，m為4且n''為3。在一特定實施例中，m為1且n''為4。在另一實施例中，m為2且n''為4。在又一實施例中，m為3且n''為4。在又另一實施例中，m為4且n''為4。在一特定實施例中，m為1且n''為5。在另一實施例中，m為2且n''為5。在又一實施例中，m為3且n''為5。在又另一實施例中，m為4且n''為5。在一特定實施例中，m為1且n''為6。在另一實施例中，m為2且n''為6。在又一實施例中，m為3且n''為6。在又另一實施例中，m為4且n''為6。In one embodiment, m is selected from 1 to 3 and n'' is selected from 0 to 10. In one embodiment, m is selected from 1 to 3 and n'' is selected from 0 to 5. In one embodiment, m is selected from 1 to 2 and n'' is selected from 0 to 3. In one embodiment, m is selected from 1 to 2 and n'' is selected from 0 to 2. In a specific embodiment, m is 1 and n'' is 0. In another embodiment, m is 2 and n'' is 0. In yet another embodiment, m is 3 and n'' is 0. In yet another embodiment, m is 4 and n'' is 0. In a specific embodiment, m is 1 and n'' is 1. In another embodiment, m is 2 and n'' is 1. In yet another embodiment, m is 3 and n'' is 1. In yet another embodiment, m is 4 and n'' is 1. In a particular embodiment, m is 1 and n'' is 2. In another embodiment, m is 2 and n'' is 2. In yet another embodiment, m is 3 and n'' is 2. In yet another embodiment, m is 4 and n'' is 2. In a particular embodiment, m is 1 and n'' is 3. In another embodiment, m is 2 and n'' is 3. In yet another embodiment, m is 3 and n'' is 3. In yet another embodiment, m is 4 and n'' is 3. In a particular embodiment, m is 1 and n'' is 4. In another embodiment, m is 2 and n'' is 4. In yet another embodiment, m is 3 and n'' is 4. In yet another embodiment, m is 4 and n'' is 4. In a particular embodiment, m is 1 and n'' is 5. In another embodiment, m is 2 and n" is 5. In yet another embodiment, m is 3 and n" is 5. In yet another embodiment, m is 4 and n" is 5. In a particular embodiment, m is 1 and n" is 6. In another embodiment, m is 2 and n" is 6. In yet another embodiment, m is 3 and n" is 6. In yet another embodiment, m is 4 and n" is 6.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(IV)，其中X為O(CH 2CH 2O) mCH 2CH 2，其中m係選自1至4，且其中X'為CH 2O(CH 2CH 2O) m '(CH 2) n ''CH 2C=O，其中n''係選自0至10且m'係選自0至4。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (IV), wherein X is O(CH 2 CH 2 O) m CH 2 CH 2 , wherein m is selected from 1 to 4, and wherein X' is CH 2 O(CH 2 CH 2 O) m ' (CH 2 ) n '' CH 2 C=O, wherein n'' is selected from 0 to 10 and m' is selected from 0 to 4.

在一實施例中，m係選自1至3，m'係選自0至4且n''係選自0至10。在一實施例中，m係選自1至2，m'係選自0至4且n''係選自0至5。在一實施例中，m係選自1至2，m'係選自0至2且n''係選自0至3。在一實施例中，m係選自1至2，m'係選自0至2且n''係選自0至1。In one embodiment, m is selected from 1 to 3, m' is selected from 0 to 4 and n'' is selected from 0 to 10. In one embodiment, m is selected from 1 to 2, m' is selected from 0 to 4 and n'' is selected from 0 to 5. In one embodiment, m is selected from 1 to 2, m' is selected from 0 to 2 and n'' is selected from 0 to 3. In one embodiment, m is selected from 1 to 2, m' is selected from 0 to 2 and n'' is selected from 0 to 1.

在一特定實施例中，m為1，m'為0且n''為0。在另一實施例中，m為1，m'為1且n''為0。在另一實施例中，m為1，m'為2且n''為0。在另一實施例中，m為1，m'為3且n''為0。In a particular embodiment, m is 1, m' is 0 and n'' is 0. In another embodiment, m is 1, m' is 1 and n'' is 0. In another embodiment, m is 1, m' is 2 and n'' is 0. In another embodiment, m is 1, m' is 3 and n'' is 0.

在另一實施例中，m為2，m'為0且n''為0。在另一實施例中，m為2，m'為1且n''為0。在另一實施例中，m為2，m'為2且n''為0。在另一實施例中，m為2，m'為3且n''為0。In another embodiment, m is 2, m' is 0 and n'' is 0. In another embodiment, m is 2, m' is 1 and n'' is 0. In another embodiment, m is 2, m' is 2 and n'' is 0. In another embodiment, m is 2, m' is 3 and n'' is 0.

在又一實施例中，m為3，m'為0且n''為0。在又一實施例中，m為3，m'為1且n''為0。在又一實施例中，m為3，m'為2且n''為0。在又一實施例中，m為3，m'為3且n''為0。In yet another embodiment, m is 3, m' is 0 and n'' is 0. In yet another embodiment, m is 3, m' is 1 and n'' is 0. In yet another embodiment, m is 3, m' is 2 and n'' is 0. In yet another embodiment, m is 3, m' is 3 and n'' is 0.

在又另一實施例中，m為4，m'為0且n''為0。在又另一實施例中，m為4，m'為1且n''為0。在又另一實施例中，m為4，m'為2且n''為0。在又另一實施例中，m為4，m'為3且n''為0。In yet another embodiment, m is 4, m' is 0 and n" is 0. In yet another embodiment, m is 4, m' is 1 and n" is 0. In yet another embodiment, m is 4, m' is 2 and n" is 0. In yet another embodiment, m is 4, m' is 3 and n" is 0.

在一特定實施例中，m為1，m'為0且n''為1。在一特定實施例中，m為1，m'為1且n''為1。在一特定實施例中，m為1，m'為2且n''為1。在一特定實施例中，m為1，m'為3且n''為1。In a specific embodiment, m is 1, m' is 0 and n'' is 1. In a specific embodiment, m is 1, m' is 1 and n'' is 1. In a specific embodiment, m is 1, m' is 2 and n'' is 1. In a specific embodiment, m is 1, m' is 3 and n'' is 1.

在另一實施例中，m為2，m'為0且n''為1。在另一實施例中，m為2，m'為1且n''為1。在另一實施例中，m為2，m'為2且n''為1。在另一實施例中，m為2，m'為3且n''為1。In another embodiment, m is 2, m' is 0 and n'' is 1. In another embodiment, m is 2, m' is 1 and n'' is 1. In another embodiment, m is 2, m' is 2 and n'' is 1. In another embodiment, m is 2, m' is 3 and n'' is 1.

在又一實施例中，m為3，m'為0且n''為1。在又一實施例中，m為3，m'為1且n''為1。在又一實施例中，m為3，m'為2且n''為1。在又一實施例中，m為3，m'為3且n''為1。In yet another embodiment, m is 3, m' is 0 and n'' is 1. In yet another embodiment, m is 3, m' is 1 and n'' is 1. In yet another embodiment, m is 3, m' is 2 and n'' is 1. In yet another embodiment, m is 3, m' is 3 and n'' is 1.

在又另一實施例中，m為4，m'為0且n''為1。在又另一實施例中，m為4，m'為1且n''為1。在又另一實施例中，m為4，m'為2且n''為1。在又另一實施例中，m為4，m'為3且n''為1。In yet another embodiment, m is 4, m' is 0 and n'' is 1. In yet another embodiment, m is 4, m' is 1 and n'' is 1. In yet another embodiment, m is 4, m' is 2 and n'' is 1. In yet another embodiment, m is 4, m' is 3 and n'' is 1.

在一特定實施例中，m為1，m'為0且n''為2。在一特定實施例中，m為1，m'為1且n''為2。在一特定實施例中，m為1，m'為2且n''為2。在一特定實施例中，m為1，m'為3且n''為2。In a specific embodiment, m is 1, m' is 0 and n'' is 2. In a specific embodiment, m is 1, m' is 1 and n'' is 2. In a specific embodiment, m is 1, m' is 2 and n'' is 2. In a specific embodiment, m is 1, m' is 3 and n'' is 2.

在另一實施例中，m為2，m'為0且n''為2。在另一實施例中，m為2，m'為1且n''為2。在另一實施例中，m為2，m'為2且n''為2。在另一實施例中，m為2，m'為3且n''為2。In another embodiment, m is 2, m' is 0 and n'' is 2. In another embodiment, m is 2, m' is 1 and n'' is 2. In another embodiment, m is 2, m' is 2 and n'' is 2. In another embodiment, m is 2, m' is 3 and n'' is 2.

在又一實施例中，m為3，m'為0且n''為2。在又一實施例中，m為3，m'為1且n''為2。在又一實施例中，m為3，m'為2且n''為2。在又一實施例中，m為3，m'為3且n''為2。In yet another embodiment, m is 3, m' is 0 and n'' is 2. In yet another embodiment, m is 3, m' is 1 and n'' is 2. In yet another embodiment, m is 3, m' is 2 and n'' is 2. In yet another embodiment, m is 3, m' is 3 and n'' is 2.

在又另一實施例中，m為4，m'為0且n''為2。在又另一實施例中，m為4，m'為1且n''為2。在又另一實施例中，m為4，m'為2且n''為2。在又另一實施例中，m為4，m'為3且n''為2。In yet another embodiment, m is 4, m' is 0 and n'' is 2. In yet another embodiment, m is 4, m' is 1 and n'' is 2. In yet another embodiment, m is 4, m' is 2 and n'' is 2. In yet another embodiment, m is 4, m' is 3 and n'' is 2.

在一特定實施例中，m為1，m'為0且n''為3。在一特定實施例中，m為1，m'為1且n''為3。在一特定實施例中，m為1，m'為2且n''為3。在一特定實施例中，m為1，m'為3且n''為3。In a specific embodiment, m is 1, m' is 0 and n'' is 3. In a specific embodiment, m is 1, m' is 1 and n'' is 3. In a specific embodiment, m is 1, m' is 2 and n'' is 3. In a specific embodiment, m is 1, m' is 3 and n'' is 3.

在另一實施例中，m為2，m'為0且n''為3。在另一實施例中，m為2，m'為1且n''為3。在另一實施例中，m為2，m'為2且n''為3。在另一實施例中，m為2，m'為3且n''為3。In another embodiment, m is 2, m' is 0 and n'' is 3. In another embodiment, m is 2, m' is 1 and n'' is 3. In another embodiment, m is 2, m' is 2 and n'' is 3. In another embodiment, m is 2, m' is 3 and n'' is 3.

在又一實施例中，m為3，m'為0且n''為3。在又一實施例中，m為3，m'為1且n''為3。在又一實施例中，m為3，m'為2且n''為3。在又一實施例中，m為3，m'為3且n''為3。In yet another embodiment, m is 3, m' is 0 and n'' is 3. In yet another embodiment, m is 3, m' is 1 and n'' is 3. In yet another embodiment, m is 3, m' is 2 and n'' is 3. In yet another embodiment, m is 3, m' is 3 and n'' is 3.

在又另一實施例中，m為4，m'為0且n''為3。在又另一實施例中，m為4，m'為1且n''為3。在又另一實施例中，m為4，m'為2且n''為3。在又另一實施例中，m為4，m'為3且n''為3。In yet another embodiment, m is 4, m' is 0 and n'' is 3. In yet another embodiment, m is 4, m' is 1 and n'' is 3. In yet another embodiment, m is 4, m' is 2 and n'' is 3. In yet another embodiment, m is 4, m' is 3 and n'' is 3.

在一特定實施例中，m為1，m'為0且n''為4。在一特定實施例中，m為1，m'為1且n''為4。在一特定實施例中，m為1，m'為2且n''為4。在一特定實施例中，m為1，m'為3且n''為4。In a specific embodiment, m is 1, m' is 0 and n'' is 4. In a specific embodiment, m is 1, m' is 1 and n'' is 4. In a specific embodiment, m is 1, m' is 2 and n'' is 4. In a specific embodiment, m is 1, m' is 3 and n'' is 4.

在另一實施例中，m為2，m'為0且n''為4。在另一實施例中，m為2，m'為1且n''為4。在另一實施例中，m為2，m'為2且n''為4。在另一實施例中，m為2，m'為3且n''為4。In another embodiment, m is 2, m' is 0 and n'' is 4. In another embodiment, m is 2, m' is 1 and n'' is 4. In another embodiment, m is 2, m' is 2 and n'' is 4. In another embodiment, m is 2, m' is 3 and n'' is 4.

在又一實施例中，m為3，m'為0且n''為4。在又一實施例中，m為3，m'為1且n''為4。在又一實施例中，m為3，m'為2且n''為4。在又一實施例中，m為3，m'為3且n''為4。In yet another embodiment, m is 3, m' is 0 and n'' is 4. In yet another embodiment, m is 3, m' is 1 and n'' is 4. In yet another embodiment, m is 3, m' is 2 and n'' is 4. In yet another embodiment, m is 3, m' is 3 and n'' is 4.

在又另一實施例中，m為4，m'為0且n''為4。在又另一實施例中，m為4，m'為1且n''為4。在又另一實施例中，m為4，m'為2且n''為4。在又另一實施例中，m為4，m'為3且n''為4。In yet another embodiment, m is 4, m' is 0 and n'' is 4. In yet another embodiment, m is 4, m' is 1 and n'' is 4. In yet another embodiment, m is 4, m' is 2 and n'' is 4. In yet another embodiment, m is 4, m' is 3 and n'' is 4.

在一特定實施例中，m為1，m'為0且n''為5。在一特定實施例中，m為1，m'為1且n''為5。在一特定實施例中，m為1，m'為2且n''為5。在一特定實施例中，m為1，m'為3且n''為5。In a specific embodiment, m is 1, m' is 0 and n'' is 5. In a specific embodiment, m is 1, m' is 1 and n'' is 5. In a specific embodiment, m is 1, m' is 2 and n'' is 5. In a specific embodiment, m is 1, m' is 3 and n'' is 5.

在另一實施例中，m為2，m'為0且n''為5。在另一實施例中，m為2，m'為1且n''為5。在另一實施例中，m為2，m'為2且n''為5。在另一實施例中，m為2，m'為3且n''為5。In another embodiment, m is 2, m' is 0 and n'' is 5. In another embodiment, m is 2, m' is 1 and n'' is 5. In another embodiment, m is 2, m' is 2 and n'' is 5. In another embodiment, m is 2, m' is 3 and n'' is 5.

在又一實施例中，m為3，m'為0且n''為5。在又一實施例中，m為3，m'為1且n''為5。在又一實施例中，m為3，m'為2且n''為5。在又一實施例中，m為3，m'為3且n''為5。In yet another embodiment, m is 3, m' is 0 and n'' is 5. In yet another embodiment, m is 3, m' is 1 and n'' is 5. In yet another embodiment, m is 3, m' is 2 and n'' is 5. In yet another embodiment, m is 3, m' is 3 and n'' is 5.

在又另一實施例中，m為4，m'為0且n''為5。在又另一實施例中，m為4，m'為1且n''為5。在又另一實施例中，m為4，m'為2且n''為5。在又另一實施例中，m為4，m'為3且n''為5。In yet another embodiment, m is 4, m' is 0 and n'' is 5. In yet another embodiment, m is 4, m' is 1 and n'' is 5. In yet another embodiment, m is 4, m' is 2 and n'' is 5. In yet another embodiment, m is 4, m' is 3 and n'' is 5.

在本發明之一極佳實施例中，使用點擊化學製備本發明之血清型38醣結合物。本發明亦係關於製備如上文所揭露之血清型38醣結合物的方法。In a preferred embodiment of the present invention, the serotype 38 saccharide conjugate of the present invention is prepared using click chemistry. The present invention also relates to a method for preparing the serotype 38 saccharide conjugate as disclosed above.

在一實施例中，點擊化學可包含三個步驟，(a)使經分離血清型38醣與碳酸衍生物及疊氮基連接子在非質子性溶劑中反應以產生經活化之疊氮基醣(醣的活化)，(b)使載體蛋白與帶有N-羥基丁二醯亞胺(NHS)部分和炔烴基之試劑反應，其中NHS部分與胺基反應以形成醯胺鍵，從而獲得炔烴官能化載體蛋白(載體蛋白的活化)，(c)藉由Cu + 1介導之疊氮化物-炔烴環加成反應使步驟(a)之經活化之疊氮基醣與步驟(b)之經活化的炔烴-載體蛋白反應以形成醣結合物。 In one embodiment, the click chemistry may comprise three steps, (a) reacting the isolated serotype 38 sugar with a carbonate derivative and an azido linker in an aprotic solvent to produce an activated azido sugar (sugar activation), (b) reacting the carrier protein with a reagent having an N-hydroxysuccinimide (NHS) moiety and an alkynyl group, wherein the NHS moiety reacts with an amine group to form an amide bond, thereby obtaining an alkynyl functionalized carrier protein (carrier protein activation), (c) reacting the carrier protein with a Cu + 1 -mediated azido-alkyne cycloaddition reaction allows the activated azido sugar of step (a) to react with the activated alkyne-carrier protein of step (b) to form a sugar conjugate.

在步驟(a)之後，醣被稱為經活化，且在本文中被稱作「經活化之醣」或「經活化之疊氮基醣」。After step (a), the carbohydrate is said to be activated and is referred to herein as "activated carbohydrate" or "activated aryl carbohydrate".

在步驟(b)之後，載體被稱為經活化，且被稱作「經活化之載體」。After step (b), the vector is said to be activated and is referred to as "activated vector".

如上文所提及，在活化(a)之前，可將醣之尺寸設定為目標分子量(MW)範圍。因此，在一實施例中，經分離之血清型38醣在用碳酸衍生物及疊氮基連接子活化之前經尺寸設定。在一實施例中，經分離之血清型38醣經尺寸設定為上文所定義之目標分子量(MW)範圍中之任一者。在一實施例中，經分離之血清型38醣在用碳酸衍生物及疊氮基連接子活化之前經尺寸設定。As mentioned above, prior to activation (a), the saccharides may be sized to a target molecular weight (MW) range. Thus, in one embodiment, the isolated serotype 38 saccharides are sized prior to activation with a carbonate derivative and an azido linker. In one embodiment, the isolated serotype 38 saccharides are sized to any of the target molecular weight (MW) ranges defined above. In one embodiment, the isolated serotype 38 saccharides are sized prior to activation with a carbonate derivative and an azido linker.

在一實施例中，該碳酸衍生物係選自由以下組成之群：1,1'-羰基二咪唑(CDI)、1,1'-羰基-二-(1,2,4-三唑) (CDT)、碳酸N,N'-二丁二醯亞胺酯(DSC)及氯甲酸N-羥基丁二醯亞胺酯。In one embodiment, the carbonic acid derivative is selected from the group consisting of 1,1'-carbonyldiimidazole (CDI), 1,1'-carbonyl-di-(1,2,4-triazole) (CDT), N,N'-disuccinimidyl carbonate (DSC) and N-hydroxysuccinimidyl chloroformate.

在一實施例中，該碳酸衍生物為1,1'-羰基二咪唑(CDI)。在另一實施例中，該碳酸衍生物為1,1'-羰基-二-(1,2,4-三唑) (CDT)。在另一實施例中，該碳酸衍生物為碳酸N,N'-二丁二醯亞胺酯(DSC)。在又另一實施例中，該碳酸衍生物為氯甲酸N-羥基丁二醯亞胺酯。In one embodiment, the carbonic acid derivative is 1,1'-carbonyldiimidazole (CDI). In another embodiment, the carbonic acid derivative is 1,1'-carbonyl-di-(1,2,4-triazole) (CDT). In another embodiment, the carbonic acid derivative is N,N'-disuccinimidyl carbonate (DSC). In yet another embodiment, the carbonic acid derivative is N-hydroxysuccinimidyl chloroformate.

在一實施例中，該碳酸衍生物為1,1'-羰基二咪唑(CDI)或1,1'-羰基-二-(1,2,4-三唑) (CDT)。在一實施例中，該碳酸衍生物為1,1'-羰基二咪唑(CDI)。較佳地，該碳酸衍生物為碳酸N,N'-二丁二醯亞胺酯(DSC)。In one embodiment, the carbonic acid derivative is 1,1'-carbonyldiimidazole (CDI) or 1,1'-carbonyl-di-(1,2,4-triazole) (CDT). In one embodiment, the carbonic acid derivative is 1,1'-carbonyldiimidazole (CDI). Preferably, the carbonic acid derivative is N,N'-disuccinimidyl carbonate (DSC).

在一實施例中，該疊氮基連接子為式(VI)之化合物， 其中X係選自由CH 2(CH 2) n、(CH 2CH 2O) mCH 2CH 2、NHCO(CH 2) n、NHCO(CH 2CH 2O) mCH 2CH 2、OCH 2(CH 2) n及O(CH 2CH 2O) mCH 2CH 2組成之群；其中n係選自1至10且m係選自1至4。 In one embodiment, the azido linker is a compound of formula (VI), wherein X is selected from the group consisting of CH2 ( CH2 ) n , ( CH2CH2O ) mCH2CH2 , NHCO ( CH2 ) n , NHCO( CH2CH2O ) mCH2CH2 , OCH2 ( CH2 ) n , and O( CH2CH2O ) mCH2CH2 ; wherein n is selected from 1-10 and m is selected from 1-4 .

在一實施例中，該疊氮基連接子為式(VI)之化合物，其中X為CH 2(CH 2) n，且n係選自1至10。在一實施例中，n係選自1至5。在一實施例中，n係選自1至4。在一實施例中，n係選自1至3。在一實施例中，n係選自1至2。在一特定實施例中，n為1。在另一實施例中，n為2。在又一實施例中，n為3。在又另一實施例中，n為4。在又另一實施例中，n為5。在又另一實施例中，n為6。在又另一實施例中，n為7。在又另一實施例中，n為8。在又另一實施例中，n為9。在又另一實施例中，n為10。 In one embodiment, the azido linker is a compound of formula (VI), wherein X is CH2 ( CH2 ) n , and n is selected from 1 to 10. In one embodiment, n is selected from 1 to 5. In one embodiment, n is selected from 1 to 4. In one embodiment, n is selected from 1 to 3. In one embodiment, n is selected from 1 to 2. In a particular embodiment, n is 1. In another embodiment, n is 2. In another embodiment, n is 3. In yet another embodiment, n is 4. In yet another embodiment, n is 5. In yet another embodiment, n is 6. In yet another embodiment, n is 7. In yet another embodiment, n is 8. In yet another embodiment, n is 9. In yet another embodiment, n is 10.

在一實施例中，該疊氮基連接子為式(VI)之化合物，其中X為(CH 2CH 2O) mCH 2CH 2，其中m係選自1至4。在一實施例中，m係選自1至3。在一實施例中，m係選自1至2。在一特定實施例中，m為1。在另一實施例中，m為2。在又一實施例中，m為3。在又另一實施例中，m為4。 In one embodiment, the azido linker is a compound of formula (VI), wherein X is ( CH2CH2O ) mCH2CH2 , wherein m is selected from 1 to 4. In one embodiment, m is selected from 1 to 3. In one embodiment, m is selected from 1 to 2. In a specific embodiment, m is 1. In another embodiment, m is 2. In yet another embodiment, m is 3. In yet another embodiment, m is 4.

在一實施例中，該疊氮基連接子為式(VI)之化合物，其中X為NHCO(CH 2) n，且n係選自1至10。在一實施例中，n係選自1至5。在一實施例中，n係選自1至4。在一實施例中，n係選自1至3。在一實施例中，n係選自1至2。在一特定實施例中，n為1。在另一實施例中，n為2。在又一實施例中，n為3。在又另一實施例中，n為4。在又另一實施例中，n為5。在又另一實施例中，n為6。在又另一實施例中，n為7。在又另一實施例中，n為8。在又另一實施例中，n為9。在又另一實施例中，n為10。 In one embodiment, the azido linker is a compound of formula (VI), wherein X is NHCO(CH 2 ) n , and n is selected from 1 to 10. In one embodiment, n is selected from 1 to 5. In one embodiment, n is selected from 1 to 4. In one embodiment, n is selected from 1 to 3. In one embodiment, n is selected from 1 to 2. In a particular embodiment, n is 1. In another embodiment, n is 2. In another embodiment, n is 3. In yet another embodiment, n is 4. In yet another embodiment, n is 5. In yet another embodiment, n is 6. In yet another embodiment, n is 7. In yet another embodiment, n is 8. In yet another embodiment, n is 9. In yet another embodiment, n is 10.

在一實施例中，該疊氮基連接子為式(VI)之化合物，其中X為NHCO(CH 2CH 2O) mCH 2CH 2，其中m係選自1至4。在一實施例中，m係選自1至3。在一實施例中，m係選自1至2。在一特定實施例中，m為1。在另一實施例中，m為2。在又一實施例中，m為3。在又另一實施例中，m為4。 In one embodiment, the azido linker is a compound of formula (VI), wherein X is NHCO(CH 2 CH 2 O) m CH 2 CH 2 , wherein m is selected from 1 to 4. In one embodiment, m is selected from 1 to 3. In one embodiment, m is selected from 1 to 2. In a specific embodiment, m is 1. In another embodiment, m is 2. In yet another embodiment, m is 3. In yet another embodiment, m is 4.

在一實施例中，該疊氮基連接子為式(VI)之化合物，其中X為OCH 2(CH 2) n，且n係選自1至10。在一實施例中，n係選自1至5。在一實施例中，n係選自1至4。在一實施例中，n係選自1至3。在一實施例中，n係選自1至2。在一特定實施例中，n為1。在另一實施例中，n為2。在又一實施例中，n為3。在又另一實施例中，n為4。在又另一實施例中，n為5。在又另一實施例中，n為6。在又另一實施例中，n為7。在又另一實施例中，n為8。在又另一實施例中，n為9。在又另一實施例中，n為10。 In one embodiment, the azido linker is a compound of formula (VI), wherein X is OCH2 ( CH2 ) n , and n is selected from 1 to 10. In one embodiment, n is selected from 1 to 5. In one embodiment, n is selected from 1 to 4. In one embodiment, n is selected from 1 to 3. In one embodiment, n is selected from 1 to 2. In a particular embodiment, n is 1. In another embodiment, n is 2. In another embodiment, n is 3. In yet another embodiment, n is 4. In yet another embodiment, n is 5. In yet another embodiment, n is 6. In yet another embodiment, n is 7. In yet another embodiment, n is 8. In yet another embodiment, n is 9. In yet another embodiment, n is 10.

在一實施例中，該疊氮基連接子為式(VI)之化合物，其中X為O(CH 2CH 2O) mCH 2CH 2，其中m係選自1至4。在一實施例中，m係選自1至3。在一實施例中，m係選自1至2。在一特定實施例中，m為1。在另一實施例中，m為2。在又一實施例中，m為3。在又另一實施例中，m為4。 In one embodiment, the azido linker is a compound of formula (VI), wherein X is O( CH2CH2O ) mCH2CH2 , wherein m is selected from 1 to 4. In one embodiment, m is selected from 1 to 3. In one embodiment, m is selected from 1 to 2. In a specific embodiment, m is 1. In another embodiment, m is 2. In yet another embodiment, m is 3. In yet another embodiment, m is 4.

在一實施例中，該疊氮基連接子為式(VII)之化合物， In one embodiment, the azido linker is a compound of formula (VII),

在一較佳實施例中，該疊氮基連接子為3-疊氮基-丙胺。In a preferred embodiment, the azido linker is 3-azido-propylamine.

在一實施例中，該帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑為帶有N-羥基丁二醯亞胺(NHS)部分及末端炔烴之試劑。In one embodiment, the reagent having an N-hydroxysuccinimide (NHS) moiety and an alkyne group is a reagent having an N-hydroxysuccinimide (NHS) moiety and a terminal alkyne group.

在一實施例中，該帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑為帶有N-羥基丁二醯亞胺(NHS)部分及環炔烴之試劑。In one embodiment, the reagent having an N-hydroxysuccinimide (NHS) moiety and an alkynyl group is a reagent having an N-hydroxysuccinimide (NHS) moiety and a cycloalkyne group.

在一實施例中，該帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑為式(VIII)之化合物， 其中X係選自由CH 2O(CH 2) nCH 2C=O及CH 2O(CH 2CH 2O) m(CH 2) nCH 2C=O組成之群，其中n係選自0至10且m係選自0至4。 In one embodiment, the reagent having an N-hydroxysuccinimide (NHS) moiety and an alkynyl group is a compound of formula (VIII), wherein X is selected from the group consisting of CH2O ( CH2 ) nCH2C = O and CH2O ( CH2CH2O ) m ( CH2 ) nCH2C = O , wherein n is selected from 0-10 and m is selected from 0-4.

在一實施例中，該帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑為式(VIII)之化合物，其中X為CH 2O(CH 2) nCH 2C=O，其中n係選自0至10。在一實施例中，n係選自0至5。在一實施例中，n係選自0至4。在一實施例中，n係選自0至3。在一實施例中，n係選自0至2。在一特定實施例中，n為0。在一特定實施例中，n為1。在另一實施例中，n為2。在又一實施例中，n為3。在又另一實施例中，n為4。在又另一實施例中，n為5。在又另一實施例中，n為6。在又另一實施例中，n為7。在又另一實施例中，n為8。在又另一實施例中，n為9。在又另一實施例中，n為10。 In one embodiment, the reagent having an N-hydroxysuccinimide (NHS) moiety and an alkynyl group is a compound of formula (VIII), wherein X is CH 2 O(CH 2 ) n CH 2 C═O, wherein n is selected from 0 to 10. In one embodiment, n is selected from 0 to 5. In one embodiment, n is selected from 0 to 4. In one embodiment, n is selected from 0 to 3. In one embodiment, n is selected from 0 to 2. In a specific embodiment, n is 0. In a specific embodiment, n is 1. In another embodiment, n is 2. In another embodiment, n is 3. In yet another embodiment, n is 4. In yet another embodiment, n is 5. In yet another embodiment, n is 6. In yet another embodiment, n is 7. In yet another embodiment, n is 8. In yet another embodiment, n is 9. In yet another embodiment, n is 10.

在一實施例中，該帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑為式(VIII)之化合物，其中X為CH 2O(CH 2CH 2O) m(CH 2) nCH 2C=O，其中n係選自0至10且m係選自0至4。在一實施例中，n係選自0至5。在一實施例中，n係選自0至4。在一實施例中，n係選自0至3。在一實施例中，n係選自0至2。在一特定實施例中，n為0。在一特定實施例中，n為1。在另一實施例中，n為2。在又一實施例中，n為3。在又另一實施例中，n為4。在又另一實施例中，n為5。在又另一實施例中，n為6。在又另一實施例中，n為7。在又另一實施例中，n為8。在又另一實施例中，n為9。在又另一實施例中，n為10。在一實施例中，m係選自0至3。在一實施例中，m係選自0至2。在一特定實施例中，m為1。在一特定實施例中，m為1。在另一實施例中，m為2。在又一實施例中，m為3。在又另一實施例中，m為4。 In one embodiment, the reagent having an N-hydroxysuccinimide (NHS) moiety and an alkynyl group is a compound of formula (VIII), wherein X is CH2O ( CH2CH2O ) m ( CH2 ) nCH2C = O , wherein n is selected from 0 to 10 and m is selected from 0 to 4. In one embodiment, n is selected from 0 to 5. In one embodiment, n is selected from 0 to 4. In one embodiment, n is selected from 0 to 3. In one embodiment, n is selected from 0 to 2. In a specific embodiment, n is 0. In a specific embodiment, n is 1. In another embodiment, n is 2. In another embodiment, n is 3. In yet another embodiment, n is 4. In yet another embodiment, n is 5. In yet another embodiment, n is 6. In yet another embodiment, n is 7. In yet another embodiment, n is 8. In yet another embodiment, n is 9. In yet another embodiment, n is 10. In one embodiment, m is selected from 0 to 3. In one embodiment, m is selected from 0 to 2. In a specific embodiment, m is 1. In a specific embodiment, m is 1. In another embodiment, m is 2. In yet another embodiment, m is 3. In yet another embodiment, m is 4.

在一實施例中，n係選自0至5且m係選自0至3。在一實施例中，n係選自0至5且m係選自0至2。In one embodiment, n is selected from 0 to 5 and m is selected from 0 to 3. In one embodiment, n is selected from 0 to 5 and m is selected from 0 to 2.

在一實施例中，n係選自0至4且m係選自0至3。在一實施例中，n係選自0至4且m係選自0至2。In one embodiment, n is selected from 0 to 4 and m is selected from 0 to 3. In one embodiment, n is selected from 0 to 4 and m is selected from 0 to 2.

在一實施例中，n係選自0至3且m係選自0至3。在一實施例中，n係選自0至3且m係選自0至2。In one embodiment, n is selected from 0 to 3 and m is selected from 0 to 3. In one embodiment, n is selected from 0 to 3 and m is selected from 0 to 2.

在一實施例中，n係選自0至2且m係選自0至3。在一實施例中，n係選自0至2且m係選自0至2。In one embodiment, n is selected from 0 to 2 and m is selected from 0 to 3. In one embodiment, n is selected from 0 to 2 and m is selected from 0 to 2.

在一實施例中，n係選自0至1且m係選自0至3。在一實施例中，n係選自0至1且m係選自0至2。In one embodiment, n is selected from 0 to 1 and m is selected from 0 to 3. In one embodiment, n is selected from 0 to 1 and m is selected from 0 to 2.

在一實施例中，n為0且m為0。在一實施例中，n為1且m為0。在一實施例中，n為2且m為0。在一實施例中，n為3且m為0。在一實施例中，n為4且m為0。在一實施例中，n為5且m為0。在一實施例中，n為6且m為0。在一實施例中，n為7且m為0。在一實施例中，n為8且m為0。在一實施例中，n為9且m為0。在一實施例中，n為10且m為0。In one embodiment, n is 0 and m is 0. In one embodiment, n is 1 and m is 0. In one embodiment, n is 2 and m is 0. In one embodiment, n is 3 and m is 0. In one embodiment, n is 4 and m is 0. In one embodiment, n is 5 and m is 0. In one embodiment, n is 6 and m is 0. In one embodiment, n is 7 and m is 0. In one embodiment, n is 8 and m is 0. In one embodiment, n is 9 and m is 0. In one embodiment, n is 10 and m is 0.

在一實施例中，n為0且m為1。在一實施例中，n為1且m為1。在一實施例中，n為2且m為1。在一實施例中，n為3且m為1。在一實施例中，n為4且m為1。在一實施例中，n為5且m為1。在一實施例中，n為6且m為1。在一實施例中，n為7且m為1。在一實施例中，n為8且m為1。在一實施例中，n為9且m為1。在一實施例中，n為10且m為1。In one embodiment, n is 0 and m is 1. In one embodiment, n is 1 and m is 1. In one embodiment, n is 2 and m is 1. In one embodiment, n is 3 and m is 1. In one embodiment, n is 4 and m is 1. In one embodiment, n is 5 and m is 1. In one embodiment, n is 6 and m is 1. In one embodiment, n is 7 and m is 1. In one embodiment, n is 8 and m is 1. In one embodiment, n is 9 and m is 1. In one embodiment, n is 10 and m is 1.

在一實施例中，n為0且m為2。在一實施例中，n為1且m為2。在一實施例中，n為2且m為2。在一實施例中，n為3且m為2。在一實施例中，n為4且m為2。在一實施例中，n為5且m為2。在一實施例中，n為6且m為2。在一實施例中，n為7且m為2。在一實施例中，n為8且m為2。在一實施例中，n為9且m為2。在一實施例中，n為10且m為2。In one embodiment, n is 0 and m is 2. In one embodiment, n is 1 and m is 2. In one embodiment, n is 2 and m is 2. In one embodiment, n is 3 and m is 2. In one embodiment, n is 4 and m is 2. In one embodiment, n is 5 and m is 2. In one embodiment, n is 6 and m is 2. In one embodiment, n is 7 and m is 2. In one embodiment, n is 8 and m is 2. In one embodiment, n is 9 and m is 2. In one embodiment, n is 10 and m is 2.

在一實施例中，n為0且m為3。在一實施例中，n為1且m為3。在一實施例中，n為2且m為3。在一實施例中，n為3且m為3。在一實施例中，n為4且m為3。在一實施例中，n為5且m為3。在一實施例中，n為6且m為3。在一實施例中，n為7且m為3。在一實施例中，n為8且m為3。在一實施例中，n為9且m為3。在一實施例中，n為10且m為3。In one embodiment, n is 0 and m is 3. In one embodiment, n is 1 and m is 3. In one embodiment, n is 2 and m is 3. In one embodiment, n is 3 and m is 3. In one embodiment, n is 4 and m is 3. In one embodiment, n is 5 and m is 3. In one embodiment, n is 6 and m is 3. In one embodiment, n is 7 and m is 3. In one embodiment, n is 8 and m is 3. In one embodiment, n is 9 and m is 3. In one embodiment, n is 10 and m is 3.

在一實施例中，n為0且m為4。在一實施例中，n為1且m為4。在一實施例中，n為2且m為4。在一實施例中，n為3且m為4。在一實施例中，n為4且m為4。在一實施例中，n為5且m為4。在一實施例中，n為6且m為4。在一實施例中，n為7且m為4。在一實施例中，n為8且m為4。在一實施例中，n為9且m為4。在一實施例中，n為10且m為4。In one embodiment, n is 0 and m is 4. In one embodiment, n is 1 and m is 4. In one embodiment, n is 2 and m is 4. In one embodiment, n is 3 and m is 4. In one embodiment, n is 4 and m is 4. In one embodiment, n is 5 and m is 4. In one embodiment, n is 6 and m is 4. In one embodiment, n is 7 and m is 4. In one embodiment, n is 8 and m is 4. In one embodiment, n is 9 and m is 4. In one embodiment, n is 10 and m is 4.

在一實施例中，該帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑為式(IX)之化合物： In one embodiment, the reagent having an N-hydroxysuccinimide (NHS) moiety and an alkynyl group is a compound of formula (IX):

在一實施例中，步驟a)包含使醣與碳酸衍生物反應，隨後使經碳酸衍生物活化之醣與疊氮基連接子在非質子性溶劑中反應以產生經活化之疊氮基醣。In one embodiment, step a) comprises reacting a sugar with a carbonic acid derivative, and then reacting the sugar activated by the carbonic acid derivative with an azido linker in an aprotic solvent to produce an activated azido sugar.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量在0.01至10莫耳當量之間的量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the sugar with a carbonic acid derivative in an amount between 0.01 and 10 molar equivalents relative to the amount of sugar present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量在0.1至10莫耳當量之間的量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the sugar with a carbonic acid derivative in an amount between 0.1 and 10 molar equivalents relative to the amount of sugar present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量在0.5至5莫耳當量之間的量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the sugar with a carbonic acid derivative in an amount between 0.5 and 5 molar equivalents relative to the amount of sugar present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量在1至5莫耳當量之間的量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the sugar with a carbonic acid derivative in an amount between 1 and 5 molar equivalents relative to the amount of sugar present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量在2至5莫耳當量之間的量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the sugar with a carbonic acid derivative in an amount between 2 and 5 molar equivalents relative to the amount of sugar present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量在5至10莫耳當量之間的量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the sugar with a carbonic acid derivative in an amount between 5 and 10 molar equivalents relative to the amount of sugar present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量在0.1至5莫耳當量之間的量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the sugar with a carbonic acid derivative in an amount between 0.1 and 5 molar equivalents relative to the amount of sugar present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量在0.5至2莫耳當量之間的量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the sugar with a carbonic acid derivative in an amount between 0.5 and 2 molar equivalents relative to the amount of sugar present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量約0.01莫耳當量之量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the carbohydrate with a carbonic acid derivative in an amount of about 0.01 molar equivalent relative to the amount of carbohydrate present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量約0.1莫耳當量之量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the carbohydrate with a carbonic acid derivative in an amount of about 0.1 molar equivalent relative to the amount of carbohydrate present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量約0.2莫耳當量之量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the carbohydrate with a carbonic acid derivative in an amount of about 0.2 molar equivalents relative to the amount of carbohydrate present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量約0.5莫耳當量之量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the carbohydrate with a carbonic acid derivative in an amount of about 0.5 molar equivalent relative to the amount of carbohydrate present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量約1莫耳當量之量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the carbohydrate with a carbonic acid derivative in an amount of about 1 molar equivalent relative to the amount of carbohydrate present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量約2莫耳當量之量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the carbohydrate with a carbonic acid derivative in an amount of about 2 molar equivalents relative to the amount of carbohydrate present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量約5莫耳當量之量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the carbohydrate with a carbonic acid derivative in an amount of about 5 molar equivalents relative to the amount of carbohydrate present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量約10莫耳當量之量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the carbohydrate with a carbonic acid derivative in an amount of about 10 molar equivalents relative to the amount of carbohydrate present in the reaction mixture.

在一實施例中，在步驟a)處，經分離醣與碳酸衍生物在非質子性溶劑中反應。In one embodiment, at step a), the separated sugar is reacted with a carbonic acid derivative in an aprotic solvent.

在一個實施例中，經分離醣與碳酸衍生物在基本上由二甲亞碸(DMSO)或二甲基甲醯胺(DMF)組成之溶液中反應。在一個實施例中，經分離醣與碳酸衍生物在基本上由二甲基甲醯胺(DMF)組成之溶液中反應。在一個實施例中，經分離醣與碳酸衍生物在基本上由二甲亞碸(DMSO)組成之溶液中反應。In one embodiment, the separated sugars and the carbonic acid derivatives are reacted in a solution consisting essentially of dimethyl sulfoxide (DMSO) or dimethylformamide (DMF). In one embodiment, the separated sugars and the carbonic acid derivatives are reacted in a solution consisting essentially of dimethylformamide (DMF). In one embodiment, the separated sugars and the carbonic acid derivatives are reacted in a solution consisting essentially of dimethyl sulfoxide (DMSO).

在一實施例中，經分離醣與碳酸衍生物在基本上由二甲基乙醯胺組成之溶液中反應。在一實施例中，經分離醣與碳酸衍生物在基本上由N-甲基-2-吡咯啶酮組成之溶液中反應。在一實施例中，經分離醣與碳酸衍生物在基本上由六甲基磷醯胺(HMPA)組成之溶液中反應。In one embodiment, the separated sugars and the carbonic acid derivatives are reacted in a solution consisting essentially of dimethylacetamide. In one embodiment, the separated sugars and the carbonic acid derivatives are reacted in a solution consisting essentially of N-methyl-2-pyrrolidone. In one embodiment, the separated sugars and the carbonic acid derivatives are reacted in a solution consisting essentially of hexamethylphosphatamide (HMPA).

在一較佳實施例中，經分離醣與碳酸衍生物在基本上由二甲亞碸(DMSO)組成之溶液中反應。In a preferred embodiment, the separated sugar is reacted with the carbonic acid derivative in a solution consisting essentially of dimethyl sulfoxide (DMSO).

在一個實施例中，經分離醣與碳酸衍生物在二甲亞碸(DMSO)或二甲基甲醯胺(DMF)中反應。在一個實施例中，經分離醣與碳酸衍生物在二甲基甲醯胺(DMF)中反應。在一個實施例中，經分離醣與碳酸衍生物在二甲亞碸(DMSO)中反應。In one embodiment, the separated sugars are reacted with carbonic acid derivatives in dimethyl sulfoxide (DMSO) or dimethyl formamide (DMF). In one embodiment, the separated sugars are reacted with carbonic acid derivatives in dimethyl formamide (DMF). In one embodiment, the separated sugars are reacted with carbonic acid derivatives in dimethyl sulfoxide (DMSO).

在一實施例中，經分離醣與碳酸衍生物在二甲基乙醯胺中反應。在一實施例中，經分離醣與碳酸衍生物在N-甲基-2-吡咯啶酮中反應。在一實施例中，經分離醣與碳酸衍生物在六甲基磷醯胺(HMPA)中反應。In one embodiment, the separated sugars are reacted with carbonic acid derivatives in dimethylacetamide. In one embodiment, the separated sugars are reacted with carbonic acid derivatives in N-methyl-2-pyrrolidone. In one embodiment, the separated sugars are reacted with carbonic acid derivatives in hexamethylphosphatamide (HMPA).

在一較佳實施例中，經分離醣與CDI在二甲亞碸(DMSO)中反應。在一實施例中，經分離醣與CDI在無水DMSO中反應。In a preferred embodiment, the separated sugars are reacted with CDI in dimethyl sulfoxide (DMSO). In one embodiment, the separated sugars are reacted with CDI in anhydrous DMSO.

已出人意料地發現，使經分離醣與CDI在水分含量約0.1%至1% (v/v)之環境中反應能夠避免副反應。因此，在一個實施例中，經分離醣與CDI在包含0.1%至1% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含0.1%至0.5% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含0.1%至0.2% (v/v)水之非質子性溶劑中反應。It has been unexpectedly discovered that reacting the separated sugars with CDI in an environment with a moisture content of about 0.1% to 1% (v/v) can avoid side reactions. Therefore, in one embodiment, the separated sugars and CDI are reacted in an aprotic solvent containing 0.1% to 1% (v/v) water. In one embodiment, the separated sugars and CDI are reacted in an aprotic solvent containing 0.1% to 0.5% (v/v) water. In one embodiment, the separated sugars and CDI are reacted in an aprotic solvent containing 0.1% to 0.2% (v/v) water.

在一個實施例中，經分離醣與CDI在包含約0.1% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含約0.2% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含約0.3% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含約0.4% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含約0.5% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含約0.6% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含約0.7% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含約0.8% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含約0.9% (v/v)水之非質子性溶劑中反應。In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing about 0.1% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing about 0.2% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing about 0.3% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing about 0.4% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing about 0.5% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing about 0.6% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing about 0.7% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing about 0.8% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing about 0.9% (v/v) water.

在一個實施例中，經分離醣與CDI在包含0.1%至1% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含0.1%至0.5% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含0.1%至0.2% (v/v)水之DMSO中反應。In one embodiment, the separated sugars are reacted with CDI in DMSO containing 0.1% to 1% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in DMSO containing 0.1% to 0.5% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in DMSO containing 0.1% to 0.2% (v/v) water.

在一個實施例中，經分離醣與CDI在包含約0.1% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含約0.2% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含約0.3% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含約0.4% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含約0.5% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含約0.6% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含約0.7% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含約0.8% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含約0.9% (v/v)水之DMSO中反應。In one embodiment, the separated sugars and CDI are reacted in DMSO containing about 0.1% (v/v) water. In one embodiment, the separated sugars and CDI are reacted in DMSO containing about 0.2% (v/v) water. In one embodiment, the separated sugars and CDI are reacted in DMSO containing about 0.3% (v/v) water. In one embodiment, the separated sugars and CDI are reacted in DMSO containing about 0.4% (v/v) water. In one embodiment, the separated sugars and CDI are reacted in DMSO containing about 0.5% (v/v) water. In one embodiment, the separated sugars and CDI are reacted in DMSO containing about 0.6% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in DMSO containing about 0.7% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in DMSO containing about 0.8% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in DMSO containing about 0.9% (v/v) water.

在一個實施例中，在添加疊氮基連接子之前藉由添加水來淬滅游離碳酸衍生物。水可使游離CDI不活化。In one embodiment, the free carbonic acid derivative is quenched by adding water before adding the azido linker. Water can inactivate the free CDI.

因此，在一實施例中，在碳酸衍生物活化之後添加水。在一實施例中，添加水以使混合物中之總水含量達到約1%至約10% (v/v)之間。在一實施例中，添加水以使混合物中之總水含量達到約1%至約5% (v/v)之間。在一實施例中，添加水以使混合物中之總水含量達到約1% (v/v)。在一實施例中，添加水以使混合物中之總水含量達到約2% (v/v)。在一實施例中，添加水以使混合物中之總水含量達到約5% (v/v)。Therefore, in one embodiment, water is added after the carbonic acid derivative is activated. In one embodiment, water is added to bring the total water content in the mixture to between about 1% and about 10% (v/v). In one embodiment, water is added to bring the total water content in the mixture to between about 1% and about 5% (v/v). In one embodiment, water is added to bring the total water content in the mixture to about 1% (v/v). In one embodiment, water is added to bring the total water content in the mixture to about 2% (v/v). In one embodiment, water is added to bring the total water content in the mixture to about 5% (v/v).

一旦醣已與碳酸衍生物反應且在最終用水淬滅碳酸衍生物之後，經碳酸衍生物活化之醣便與疊氮基連接子反應。Once the sugar has been reacted with the carbonate derivative and after the carbonate derivative is finally quenched with water, the carbonate-activated sugar is reacted with the azido linker.

在一個實施例中，步驟a)進一步包含使經碳酸衍生物活化之醣與相對於經活化醣之多醣重複單元的量(RU之莫耳當量)在0.01至10莫耳當量之間的量的疊氮基連接子反應。In one embodiment, step a) further comprises reacting the carbonate-activated sugar with an azido linker in an amount between 0.01 and 10 molar equivalents relative to the amount of polysaccharide repeating units (molar equivalents of RU) of the activated sugar.

在一個實施例中，步驟a)進一步包含使經碳酸衍生物活化之醣與相對於經活化醣之多醣重複單元的量在0.1至5莫耳當量之間的量的疊氮基連接子反應。In one embodiment, step a) further comprises reacting the carbonate-activated carbohydrate with an azido linker in an amount between 0.1 and 5 molar equivalents relative to the amount of the polysaccharide repeating unit of the activated carbohydrate.

在一個實施例中，步驟a)進一步包含使經碳酸衍生物活化之醣與相對於經活化醣之多醣重複單元的量在0.5至2莫耳當量之間的量的疊氮基連接子反應。In one embodiment, step a) further comprises reacting the carbonate-activated carbohydrate with an azido linker in an amount between 0.5 and 2 molar equivalents relative to the amount of polysaccharide repeating units of the activated carbohydrate.

在一個實施例中，步驟a)進一步包含使經碳酸衍生物活化之醣與相對於經活化醣之多醣重複單元的量在1至5莫耳當量之間的量的疊氮基連接子反應。In one embodiment, step a) further comprises reacting the carbonate-activated carbohydrate with an azido linker in an amount between 1 and 5 molar equivalents relative to the amount of the polysaccharide repeating unit of the activated carbohydrate.

在上述實施例中，該碳酸衍生物可為CDI。在另一實施例中，該碳酸衍生物為CDT。在一較佳實施例中，該碳酸衍生物為碳酸N,N'-二丁二醯亞胺酯(DSC)。In the above embodiment, the carbonic acid derivative may be CDI. In another embodiment, the carbonic acid derivative is CDT. In a preferred embodiment, the carbonic acid derivative is N,N'-disuccinimidyl carbonate (DSC).

在一個實施例中，步驟a)之後經活化醣之活化程度在1.0至100%之間。疊氮基醣之活化程度定義為與疊氮基連接子連接之重複單元的百分比。In one embodiment, the degree of activation of the activated sugar after step a) is between 1.0 and 100%. The degree of activation of the azido sugar is defined as the percentage of repeat units linked to the azido linker.

在一個實施例中，步驟a)之後經活化醣之活化程度在5至70%之間。在另一實施例中，步驟a)之後經活化醣之活化程度在5至50%之間。In one embodiment, the degree of activation of the activated sugar after step a) is between 5 and 70%. In another embodiment, the degree of activation of the activated sugar after step a) is between 5 and 50%.

在一較佳實施例中，步驟a)之後經活化醣之活化程度在15至50%之間。In a preferred embodiment, the activation degree of the activated sugar after step a) is between 15 and 50%.

在另一實施例中，步驟a)之後經活化醣之活化程度在10至40%之間。In another embodiment, the activation degree of the activated sugar after step a) is between 10 and 40%.

在另一實施例中，步驟a)之後經活化醣之活化程度在5至15%之間。In another embodiment, the activation degree of the activated sugar after step a) is between 5 and 15%.

在另一實施例中，步驟a)之後經活化醣之活化程度在15至35%之間。In another embodiment, the activation degree of the activated sugar after step a) is between 15 and 35%.

在一較佳實施例中，步驟a)之後經活化醣之活化程度在15至50%之間。In a preferred embodiment, the activation degree of the activated sugar after step a) is between 15 and 50%.

在一實施例中，步驟a)之後經活化醣之活化程度為約25%。In one embodiment, the degree of activation of the activated sugar after step a) is about 25%.

在一實施例中，步驟a)之後經活化醣之活化程度為約30%。In one embodiment, the degree of activation of the activated sugar after step a) is about 30%.

在一個實施例中，步驟b)包含使載體蛋白與相對於載體上之離胺酸0.1至10莫耳當量之量的帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑反應。In one embodiment, step b) comprises reacting the carrier protein with a reagent having an N-hydroxysuccinimide (NHS) moiety and an alkyne group in an amount of 0.1 to 10 molar equivalents relative to the lysine on the carrier.

在一個實施例中，步驟b)包含使載體蛋白與相對於載體上之離胺酸0.5至10莫耳當量之量的帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑反應。In one embodiment, step b) comprises reacting the carrier protein with a reagent having an N-hydroxysuccinimide (NHS) moiety and an alkyne group in an amount of 0.5 to 10 molar equivalents relative to the lysine on the carrier.

在一個實施例中，步驟b)包含使載體蛋白與相對於載體上之離胺酸1至5莫耳當量之量的帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑反應。In one embodiment, step b) comprises reacting the carrier protein with 1 to 5 molar equivalents of a reagent having an N-hydroxysuccinimide (NHS) moiety and an alkyne group relative to the lysine on the carrier.

在一個實施例中，步驟b)包含使載體蛋白與相對於載體上之離胺酸2至5莫耳當量之量的帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑反應。In one embodiment, step b) comprises reacting the carrier protein with 2 to 5 molar equivalents of a reagent having an N-hydroxysuccinimide (NHS) moiety and an alkyne group relative to the lysine on the carrier.

在一個實施例中，步驟b)包含使載體蛋白與相對於載體上之離胺酸5至10莫耳當量之量的帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑反應。In one embodiment, step b) comprises reacting the carrier protein with a reagent having an N-hydroxysuccinimide (NHS) moiety and an alkyne group in an amount of 5 to 10 molar equivalents relative to the lysine on the carrier.

在一個實施例中，步驟b)包含使載體蛋白與相對於載體上之離胺酸1至5莫耳當量之量的帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑反應。In one embodiment, step b) comprises reacting the carrier protein with 1 to 5 molar equivalents of a reagent having an N-hydroxysuccinimide (NHS) moiety and an alkyne group relative to the lysine on the carrier.

在一個實施例中，步驟b)包含使載體蛋白與相對於載體上之離胺酸約10莫耳當量之量的帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑反應。In one embodiment, step b) comprises reacting the carrier protein with a reagent having an N-hydroxysuccinimide (NHS) moiety and an alkyne group in an amount of about 10 molar equivalents relative to the lysine on the carrier.

在一個實施例中，步驟b)包含使載體蛋白與相對於載體上之離胺酸約5莫耳當量之量的帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑反應。In one embodiment, step b) comprises reacting the carrier protein with a reagent having an N-hydroxysuccinimide (NHS) moiety and an alkyne group in an amount of about 5 molar equivalents relative to the lysine on the carrier.

在一個實施例中，步驟b)包含使載體蛋白與相對於載體上之離胺酸約2莫耳當量之量的帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑反應。In one embodiment, step b) comprises reacting the carrier protein with a reagent having an N-hydroxysuccinimide (NHS) moiety and an alkyne group in an amount of about 2 molar equivalents relative to the lysine on the carrier.

在一個實施例中，步驟b)包含使載體蛋白與相對於載體上之離胺酸約1莫耳當量之量的帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑反應。In one embodiment, step b) comprises reacting the carrier protein with a reagent having an N-hydroxysuccinimide (NHS) moiety and an alkyne group in an amount of about 1 molar equivalent relative to the lysine on the carrier.

在一個實施例中，步驟b)包含使載體蛋白與相對於載體上之離胺酸約0.5莫耳當量之量的帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑反應。In one embodiment, step b) comprises reacting the carrier protein with a reagent having an N-hydroxysuccinimide (NHS) moiety and an alkyne group in an amount of about 0.5 molar equivalent relative to the lysine on the carrier.

在一個實施例中，步驟b)包含使載體蛋白與相對於載體上之離胺酸約0.1莫耳當量之量的帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑反應。In one embodiment, step b) comprises reacting the carrier protein with a reagent having an N-hydroxysuccinimide (NHS) moiety and an alkyne group in an amount of about 0.1 molar equivalent relative to the lysine on the carrier.

在一個實施例中，步驟b)之後經活化載體之活化程度在1與50之間。經活化載體之活化程度定義為載體蛋白中變得與帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑連接的離胺酸殘基之數目。In one embodiment, the degree of activation of the activated carrier after step b) is between 1 and 50. The degree of activation of the activated carrier is defined as the number of lysine residues in the carrier protein that become linked to the reagent with an N-hydroxysuccinimide (NHS) moiety and an alkyne group.

在一實施例中，載體蛋白為CRM 197，其含有39個離胺酸殘基。在該實施例中，步驟b)之後經活化載體之活化程度可在1至30之間。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度在5至20之間。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度在9至18之間。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度在8至11之間。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度在15至20之間。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約5。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約6。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約7。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約8。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約9。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約10。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約11。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約12。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約13。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約14。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約15。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約16。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約17。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約18。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約19。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約20。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約21。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約22。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約23。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約24。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約25。 In one embodiment, the carrier protein is CRM 197 , which contains 39 lysine residues. In this embodiment, the degree of activation of the activated carrier after step b) may be between 1 and 30. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is between 5 and 20. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is between 9 and 18. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is between 8 and 11. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is between 15 and 20. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 5. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 6. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 7. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 8. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 9. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 10. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 11. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 12. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 13. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 14. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 15. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 16. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 17. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 18. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 19. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 20. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 21. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 22. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 23. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 24. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 25.

在一實施例中，載體蛋白為SCP或其片段。在該實施例中，步驟b)之後經活化載體之活化程度可在1至50之間。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度在5至50之間。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度在7至45之間。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度在5至25之間。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度在10至25之間。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度在17至22之間。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約5。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約7。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約10。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約13。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約15。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約20。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約26。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約30。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約35。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約37。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約40。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約45。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約50。In one embodiment, the carrier protein is SCP or a fragment thereof. In this embodiment, the degree of activation of the activated carrier after step b) may be between 1 and 50. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is between 5 and 50. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is between 7 and 45. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is between 5 and 25. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is between 10 and 25. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is between 17 and 22. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 5. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 7. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 10. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 13. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 15. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 20. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 26. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 30. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 35. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 37. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 40. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 45. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 50.

在一實施例中，載體蛋白為TT或其片段。在該實施例中，步驟b)之後經活化載體之活化程度可在1至30之間。在另一實施例中，步驟b)之後經活化載體(TT)之活化程度在5至25之間。在另一實施例中，步驟b)之後經活化載體(TT)之活化程度在7至25之間。在另一實施例中，步驟b)之後經活化載體(TT)之活化程度在10至20之間。在另一實施例中，步驟b)之後經活化載體(TT)之活化程度為約5。在另一實施例中，步驟b)之後經活化載體(TT)之活化程度為約7。在另一實施例中，步驟b)之後經活化載體(TT)之活化程度為約10。在另一實施例中，步驟b)之後經活化載體(TT)之活化程度為約12。在另一實施例中，步驟b)之後經活化載體(TT)之活化程度為約15。在另一實施例中，步驟b)之後經活化載體(TT)之活化程度為約20。在另一實施例中，步驟b)之後經活化載體(TT)之活化程度為約25。在另一實施例中，步驟b)之後經活化載體(TT)之活化程度為約30。In one embodiment, the carrier protein is TT or a fragment thereof. In this embodiment, the degree of activation of the activated carrier after step b) may be between 1 and 30. In another embodiment, the degree of activation of the activated carrier (TT) after step b) is between 5 and 25. In another embodiment, the degree of activation of the activated carrier (TT) after step b) is between 7 and 25. In another embodiment, the degree of activation of the activated carrier (TT) after step b) is between 10 and 20. In another embodiment, the degree of activation of the activated carrier (TT) after step b) is about 5. In another embodiment, the degree of activation of the activated carrier (TT) after step b) is about 7. In another embodiment, the degree of activation of the activated carrier (TT) after step b) is about 10. In another embodiment, the degree of activation of the activated carrier (TT) after step b) is about 12. In another embodiment, the degree of activation of the activated carrier (TT) after step b) is about 15. In another embodiment, the degree of activation of the activated carrier (TT) after step b) is about 20. In another embodiment, the degree of activation of the activated carrier (TT) after step b) is about 25. In another embodiment, the degree of activation of the activated carrier (TT) after step b) is about 30.

在一實施例中，結合反應c)係在水性緩衝液中進行。在一實施例中，結合反應c)係在水性緩衝液中在作為催化劑之銅(I)存在下進行。在一實施例中，結合反應c)係在水性緩衝液中在氧化劑及作為催化劑之銅(I)存在下進行。在一較佳實施例中，結合反應c)係在水性緩衝液中在作為催化劑之銅(I)及作為氧化劑之抗壞血酸鹽存在下進行。在一實施例中，可進一步添加THPTA (參(3-羥丙基三唑基甲基)胺)及胺基胍以防止蛋白質發生副反應。因此，在一較佳實施例中，結合反應c)係在水性緩衝液中在作為催化劑之銅(I)及作為氧化劑之抗壞血酸鹽存在下進行，其中反應混合物進一步包含THPTA (參(3-羥丙基三唑基甲基)胺)及胺基胍。In one embodiment, the binding reaction c) is carried out in an aqueous buffer. In one embodiment, the binding reaction c) is carried out in an aqueous buffer in the presence of copper (I) as a catalyst. In one embodiment, the binding reaction c) is carried out in an aqueous buffer in the presence of an oxidant and copper (I) as a catalyst. In a preferred embodiment, the binding reaction c) is carried out in an aqueous buffer in the presence of copper (I) as a catalyst and ascorbate as an oxidant. In one embodiment, THPTA (tris(3-hydroxypropyltriazolylmethyl)amine) and aminoguanidine may be further added to prevent side reactions of the protein. Therefore, in a preferred embodiment, the combination reaction c) is carried out in an aqueous buffer in the presence of copper (I) as a catalyst and ascorbate as an oxidant, wherein the reaction mixture further comprises THPTA (tris(3-hydroxypropyltriazolylmethyl)amine) and aminoguanidine.

在一實施例中，在步驟c)處經活化疊氮基醣與經活化炔烴-載體之初始輸入比率(重量/重量)在0.1與3之間。在一實施例中，在步驟c)處經活化疊氮基醣與經活化炔烴-載體之初始輸入比率(重量/重量)在0.5與2之間。在一實施例中，在步驟c)處經活化疊氮基醣與經活化炔烴-載體之初始輸入比率(重量/重量)在0.6與1.5之間。在一較佳實施例中，在步驟c)處經活化疊氮基醣與經活化炔烴-載體之初始輸入比率(重量/重量)在0.8與1之間。在一實施例中，在步驟c)處經活化疊氮基醣與經活化炔烴-載體之初始輸入比率(重量/重量)為約0.5。在一實施例中，在步驟c)處經活化疊氮基醣與經活化炔烴-載體之初始輸入比率(重量/重量)為約0.6。在一實施例中，在步驟c)處經活化疊氮基醣與經活化炔烴-載體之初始輸入比率(重量/重量)為約0.7。在一實施例中，在步驟c)處經活化疊氮基醣與經活化炔烴-載體之初始輸入比率(重量/重量)為約0.8。在一實施例中，在步驟c)處經活化疊氮基醣與經活化炔烴-載體之初始輸入比率(重量/重量)為約0.9。在一實施例中，在步驟c)處經活化疊氮基醣與經活化炔烴-載體之初始輸入比率(重量/重量)為約1。在一實施例中，在步驟c)處經活化疊氮基醣與經活化炔烴-載體之初始輸入比率(重量/重量)為約1.1。在一實施例中，在步驟c)處經活化疊氮基醣與經活化炔烴-載體之初始輸入比率(重量/重量)為約1.2。在一實施例中，在步驟c)處經活化疊氮基醣與經活化炔烴-載體之初始輸入比率(重量/重量)為約1.3。在一實施例中，在步驟c)處經活化疊氮基醣與經活化炔烴-載體之初始輸入比率(重量/重量)為約1.4。在一實施例中，在步驟c)處經活化疊氮基醣與經活化炔烴-載體之初始輸入比率(重量/重量)為約1.5。在一實施例中，在步驟c)處經活化疊氮基醣與經活化炔烴-載體之初始輸入比率(重量/重量)為約1.6。在一實施例中，在步驟c)處經活化疊氮基醣與經活化炔烴-載體之初始輸入比率(重量/重量)為約1.7。在一實施例中，在步驟c)處經活化疊氮基醣與經活化炔烴-載體之初始輸入比率(重量/重量)為約1.8。在一實施例中，在步驟c)處經活化疊氮基醣與經活化炔烴-載體之初始輸入比率(重量/重量)為約1.9。在一實施例中，在步驟c)處經活化疊氮基醣與經活化炔烴-載體之初始輸入比率(重量/重量)為約2。In one embodiment, the initial input ratio (weight/weight) of activated azidosugar to activated alkyne-carrier at step c) is between 0.1 and 3. In one embodiment, the initial input ratio (weight/weight) of activated azidosugar to activated alkyne-carrier at step c) is between 0.5 and 2. In one embodiment, the initial input ratio (weight/weight) of activated azidosugar to activated alkyne-carrier at step c) is between 0.6 and 1.5. In a preferred embodiment, the initial input ratio (weight/weight) of activated azidosugar to activated alkyne-carrier at step c) is between 0.8 and 1. In one embodiment, the initial input ratio (weight/weight) of activated azidosugar to activated alkyne-carrier at step c) is about 0.5. In one embodiment, the initial input ratio (weight/weight) of activated azidosugar to activated alkyne-carrier at step c) is about 0.6. In one embodiment, the initial input ratio (weight/weight) of activated azidosugar to activated alkyne-carrier at step c) is about 0.7. In one embodiment, the initial input ratio (weight/weight) of activated azidosugar to activated alkyne-carrier at step c) is about 0.8. In one embodiment, the initial input ratio (weight/weight) of activated azidosugar to activated alkyne-carrier at step c) is about 0.9. In one embodiment, the initial input ratio (weight/weight) of activated azidosugar to activated alkyne-carrier at step c) is about 1. In one embodiment, the initial input ratio (weight/weight) of activated azidosugar to activated alkyne-carrier at step c) is about 1.1. In one embodiment, the initial input ratio (weight/weight) of activated azidosugar to activated alkyne-carrier at step c) is about 1.2. In one embodiment, the initial input ratio (weight/weight) of activated azidosugar to activated alkyne-carrier at step c) is about 1.3. In one embodiment, the initial input ratio (weight/weight) of activated azidosugar to activated alkyne-carrier at step c) is about 1.4. In one embodiment, the initial input ratio (weight/weight) of activated azidosugar to activated alkyne-carrier at step c) is about 1.5. In one embodiment, the initial input ratio (weight/weight) of activated azidosugar to activated alkyne-carrier at step c) is about 1.6. In one embodiment, the initial input ratio (weight/weight) of activated azidosugar to activated alkyne-carrier at step c) is about 1.7. In one embodiment, the initial input ratio (weight/weight) of activated azidosugar to activated alkyne-carrier at step c) is about 1.8. In one embodiment, the initial input ratio (weight/weight) of activated azidosugar to activated alkyne-carrier at step c) is about 1.9. In one embodiment, the initial input ratio (weight/weight) of activated azidosugar to activated alkyne-carrier at step c) is about 2.

在單擊結合反應之後，結合物中可保留有未反應之疊氮基，可使用適合的疊氮基封端劑對此等未反應之疊氮基進行封端。因此，在一實施例中，在步驟c)之後，使用適合的疊氮基封端劑對結合物中未反應之疊氮基進行封端。在一個實施例中，此疊氮基封端劑為帶有炔烴基之試劑。在一個實施例中，此疊氮基封端劑為帶有末端炔烴之試劑。在一個實施例中，此疊氮基封端劑為帶有環炔烴之試劑。After the single-stranded conjugation reaction, unreacted azido groups may remain in the conjugate, and suitable azido group capping agents may be used to cap such unreacted azido groups. Therefore, in one embodiment, after step c), suitable azido group capping agents are used to cap the unreacted azido groups in the conjugate. In one embodiment, the azido group capping agent is a reagent with an alkynyl group. In one embodiment, the azido group capping agent is a reagent with a terminal alkynyl group. In one embodiment, the azido group capping agent is a reagent with a cycloalkynyl group.

在一實施例中，該疊氮基封端劑為式(X)之化合物， 其中X為(CH 2) n，其中n係選自1至15。 In one embodiment, the azido-capping agent is a compound of formula (X), wherein X is (CH 2 ) n , wherein n is selected from 1-15.

在一個實施例中，此疊氮基封端劑為炔丙醇。In one embodiment, the azido-capping agent is propargyl alcohol.

因此，在一實施例中，在步驟c)之後，該方法進一步包含用疊氮基封端劑對結合物中保留之未反應疊氮基進行封端的步驟。Therefore, in one embodiment, after step c), the method further comprises the step of capping the unreacted azido groups remaining in the conjugate with an azido capping agent.

在一實施例中，未反應疊氮基之封端係用相對於經活化醣之多醣重複單元的量在0.05至20莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted azido groups is performed with a capping agent in an amount between 0.05 and 20 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應疊氮基之封端係用相對於經活化醣之多醣重複單元的量在0.1至15莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted azido groups is performed with a capping agent in an amount between 0.1 and 15 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應疊氮基之封端係用相對於經活化醣之多醣重複單元的量在0.5至10莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted azido groups is performed with a capping agent in an amount between 0.5 and 10 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應疊氮基之封端係用相對於經活化醣之多醣重複單元的量在0.5至5莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted azido groups is performed with an amount of the capping agent between 0.5 and 5 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應疊氮基之封端係用相對於經活化醣之多醣重複單元的量在0.5至2莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted azido groups is performed with a capping agent in an amount between 0.5 and 2 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應疊氮基之封端係用相對於經活化醣之多醣重複單元的量在0.5至1莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted azido groups is performed with a capping agent in an amount between 0.5 and 1 molar equivalent relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應疊氮基之封端係用相對於經活化醣之多醣重複單元的量在1至2莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted azido groups is performed with an amount of the capping agent between 1 and 2 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應疊氮基之封端係用相對於經活化醣之多醣重複單元的量在0.75至1.5莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted azido groups is performed with an amount of the capping agent between 0.75 and 1.5 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應疊氮基之封端係用相對於經活化醣之多醣重複單元的量約1莫耳當量之量的封端劑進行。In one embodiment, the capping of unreacted azido groups is performed using a capping agent in an amount of about 1 molar equivalent relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應疊氮基之封端係用相對於經活化醣之多醣重複單元的量約1.5莫耳當量之量的封端劑進行。In one embodiment, the capping of unreacted azido groups is performed using a capping agent in an amount of about 1.5 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應疊氮基之封端係用相對於經活化醣之多醣重複單元的量約0.5莫耳當量之量的封端劑進行。In one embodiment, the capping of unreacted azido groups is performed using a capping agent in an amount of about 0.5 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應疊氮基之封端係用相對於經活化醣之多醣重複單元的量約2莫耳當量之量的封端劑進行。In one embodiment, the capping of unreacted azido groups is performed using a capping agent in an amount of about 2 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在單擊結合反應之後，未反應之炔烴基可仍存在於結合物中，可使用適合的炔烴基封端劑對此等未反應之炔烴基進行封端。在一個實施例中，此炔烴基封端劑為帶有疊氮基之試劑。After the single-strand conjugation reaction, unreacted alkynyl groups may still exist in the conjugate, and these unreacted alkynyl groups can be capped using a suitable alkynyl capping agent. In one embodiment, the alkynyl capping agent is a reagent with an azido group.

在一實施例中，該炔烴基封端劑為式(XI)之化合物， 其中X為(CH 2) n，其中n係選自1至15。 In one embodiment, the alkynyl end-capping agent is a compound of formula (XI), wherein X is (CH 2 ) n , wherein n is selected from 1-15.

在一個實施例中，此炔烴基封端劑為3-疊氮基-1-丙醇。In one embodiment, the alkynyl end-capping agent is 3-azido-1-propanol.

因此，在一實施例中，在步驟c)之後，該方法進一步包含用炔烴基封端劑對結合物中保留之未反應炔烴基進行封端的步驟。Therefore, in one embodiment, after step c), the method further comprises the step of capping the unreacted alkynyl groups remaining in the conjugate with an alkynyl capping agent.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量在0.05至20莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed with an amount of the capping agent between 0.05 and 20 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量在0.1至15莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed with an amount of the capping agent between 0.1 and 15 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量在0.5至10莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed with an amount of the capping agent between 0.5 and 10 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量在0.5至5莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed with an amount of the capping agent between 0.5 and 5 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量在0.5至2莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed with an amount of the capping agent between 0.5 and 2 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量在0.5至1莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed with an amount of the capping agent between 0.5 and 1 molar equivalent relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量在1至5莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed with an amount of the capping agent between 1 and 5 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量在1至2莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed with an amount of the capping agent between 1 and 2 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量在1.5至2.5莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed with an amount of the capping agent between 1.5 and 2.5 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量約0.5莫耳當量之量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed using a capping agent in an amount of about 0.5 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量約1莫耳當量之量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed using a capping agent in an amount of about 1 molar equivalent relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量約1.5莫耳當量之量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed using an amount of the capping agent in an amount of about 1.5 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量約2莫耳當量之量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed using a capping agent in an amount of about 2 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量約2.5莫耳當量之量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed using an amount of the capping agent in an amount of about 2.5 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量約5莫耳當量之量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed using a capping agent in an amount of about 5 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在與載體蛋白結合之後，可藉由技術人員已知之多種技術來純化(就醣-蛋白結合物之量而言富集)醣結合物。此等技術包括透析、濃縮/透濾操作、切向流過濾沉澱/溶離、管柱層析(DEAE或疏水性相互作用層析)及深層過濾。因此，在一個實施例中，用於產生本發明之醣結合物的方法包含在產生醣結合物之後對其進行純化的步驟。After conjugation to the carrier protein, the glycoconjugate can be purified (enriched in terms of the amount of glycoconjugate) by a variety of techniques known to those skilled in the art. These techniques include dialysis, concentration/filtration procedures, tangential flow filtration precipitation/elution, column chromatography (DEAE or hydrophobic interaction chromatography) and deep filtration. Therefore, in one embodiment, the method for producing the glycoconjugate of the present invention includes a step of purifying the glycoconjugate after it is produced.

在一態樣中，本發明提供一種根據本文所揭露之方法中之任一者產生的血清型38醣結合物。In one aspect, the invention provides a serotype 38 saccharide conjugate produced according to any of the methods disclosed herein.

替代點擊化學在本發明之一個實施例中，本發明之血清型38醣結合物藉由如例如申請案第PCT/IB2024/051122號(於2024年2月07日申請)中所揭露之替代點選化學方法製備。 Substitution click chemistry In one embodiment of the present invention, the serotype 38 carbohydrate conjugate of the present invention is prepared by a substitution click chemistry method as disclosed in, for example, application No. PCT/IB2024/051122 (filed on February 7, 2024).

因此，在一實施例中，本發明之血清型38醣結合物包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(XII)： ， 其中X係選自由CH 2(CH 2) n '、(CH 2CH 2O) mCH 2CH 2、NHCO(CH 2) n '、NHCO(CH 2CH 2O) mCH 2CH 2、OCH 2(CH 2) n '及O(CH 2CH 2O) mCH 2CH 2組成之群；其中n'係選自0至10且m係選自1至4， 其中X'係選自由CH 2(CH 2) n ''、CH 2O(CH 2) n ''CH 2、CH 2O(CH 2CH 2O) m '(CH 2) n ''CH 2組成之群，其中n''係選自0至10且m'係選自0至4且其中方括號中之結構表示該血清型38醣之重複單元，且其中n表示重複單元之數目。 Therefore, in one embodiment, the serotype 38 saccharide conjugate of the present invention comprises a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and has the general formula (XII): wherein X is selected from the group consisting of CH2 ( CH2 ) n ' , ( CH2CH2O ) mCH2CH2 , NHCO( CH2 ) n ' , NHCO( CH2CH2O ) mCH2CH2 , OCH2 ( CH2 ) n ' , and O( CH2CH2O ) mCH2CH2 ; wherein n' is selected from 0 to 10 and m is selected from 1 to 4, wherein X' is selected from the group consisting of CH2 ( CH2 ) n '' , CH2O ( CH2 ) n''CH2 , CH2O (CH2CH2O)m ' , ( CH2 ) n''CH 2 , wherein n'' is selected from 0 to 10 and m' is selected from 0 to 4 and wherein the structure in square brackets represents the repeating unit of the serotype 38 carbohydrate, and wherein n represents the number of repeating units.

在一特定態樣中，本發明係關於一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(XII)，其中X為CH 2(CH 2) n '，其中n'為0，且其中X'為CH 2(CH 2) n ''，其中n''為0。 In a specific embodiment, the present invention relates to a serotype 38 saccharide conjugate comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (XII), wherein X is CH2 ( CH2 ) n ' , wherein n' is 0, and wherein X' is CH2 ( CH2 ) n '' , wherein n'' is 0.

在一特定態樣中，本發明係關於一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(XIII)， 其中方括號中之結構表示該血清型38醣之重複單元，且其中n表示重複單元之數目。 In a specific embodiment, the present invention relates to a serotype 38 saccharide conjugate, which comprises a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and has the general formula (XIII), Wherein the structure in square brackets represents the repeating unit of the serotype 38 carbohydrate, and wherein n represents the number of repeating units.

式(XII)及(XIII)為本發明之醣結合物的示意性表示。不應理解為醣之每一重複單元(方括號中之結構)處均存在連接。實情為，大部分醣重複單元保持未經修飾，且少數醣重複單元處存在載體蛋白與醣之共價連接。另外，個別載體蛋白(CP)分子可連接至多於一個醣分子，且個體醣分子可連接至多於一個個別載體蛋白(CP)分子。方括號中之結構表示血清型38醣之重複單元。Formulas (XII) and (XIII) are schematic representations of the carbohydrate conjugates of the present invention. It should not be understood that there is a linkage at every repeat unit of the carbohydrate (structure in square brackets). In fact, most carbohydrate repeat units remain unmodified, and a few carbohydrate repeat units have a covalent linkage of the carrier protein to the carbohydrate. In addition, an individual carrier protein (CP) molecule can be linked to more than one carbohydrate molecule, and an individual carbohydrate molecule can be linked to more than one individual carrier protein (CP) molecule. The structure in square brackets represents a repeat unit of a serotype 38 carbohydrate.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(XII)，其中X為CH 2(CH 2) n '，其中n'係選自0至10，且其中X'為CH 2(CH 2) n ''，其中n''係選自0至10。在一實施例中，n'係選自0至5且n''係選自0至10。在一實施例中，n'係選自0至5且n''係選自0至5。在一實施例中，n'係選自0至3且n''係選自0至3。在一實施例中，n'係選自0至2且n''係選自0至2。在一特定實施例中，n'為0且n''為0。在一特定實施例中，n'為1且n''為0。在另一實施例中，n'為2且n''為0。在又一實施例中，n'為3且n''為0。在又另一實施例中，n'為4且n''為0。在又另一實施例中，n'為5且n''為0。在又另一實施例中，n'為6且n''為0。在一特定實施例中，n'為0且n''為1。在一特定實施例中，n'為1且n''為1。在另一實施例中，n'為2且n''為1。在又一實施例中，n'為3且n''為1。在又另一實施例中，n'為4且n''為1。在又另一實施例中，n'為5且n''為1。在又另一實施例中，n'為6且n''為1。在一特定實施例中，n'為0且n''為2。在一特定實施例中，n'為1且n''為2。在另一實施例中，n'為2且n''為2。在又一實施例中，n'為3且n''為2。在又另一實施例中，n'為4且n''為2。在又另一實施例中，n'為5且n''為2。在又另一實施例中，n'為6且n''為2。在一特定實施例中，n'為0且n''為3。在一特定實施例中，n'為1且n''為3。在另一實施例中，n'為2且n''為3。在又一實施例中，n'為3且n''為3。在又另一實施例中，n'為4且n''為3。在又另一實施例中，n'為5且n''為3。在又另一實施例中，n'為6且n''為3。在一特定實施例中，n'為0且n''為4。在一特定實施例中，n'為1且n''為4。在另一實施例中，n'為2且n''為4。在又一實施例中，n'為3且n''為4。在又另一實施例中，n'為4且n''為4。在又另一實施例中，n'為5且n''為4。在又另一實施例中，n'為6且n''為4。在一特定實施例中，n'為0且n''為5。在一特定實施例中，n'為1且n''為5。在另一實施例中，n'為2且n''為5。在又一實施例中，n'為3且n''為5。在又另一實施例中，n'為4且n''為5。在又另一實施例中，n'為5且n''為5。在又另一實施例中，n'為6且n''為5。在一特定實施例中，n'為0且n''為6。在一特定實施例中，n'為1且n''為6。在另一實施例中，n'為2且n''為6。在又一實施例中，n'為3且n''為6。在又另一實施例中，n'為4且n''為6。在又另一實施例中，n'為5且n''為6。在又另一實施例中，n'為6且n''為6。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (XII), wherein X is CH 2 (CH 2 ) n ' , wherein n' is selected from 0 to 10, and wherein X' is CH 2 (CH 2 ) n '' , wherein n'' is selected from 0 to 10. In one embodiment, n' is selected from 0 to 5 and n'' is selected from 0 to 10. In one embodiment, n' is selected from 0 to 5 and n'' is selected from 0 to 5. In one embodiment, n' is selected from 0 to 3 and n'' is selected from 0 to 3. In one embodiment, n' is selected from 0 to 2 and n'' is selected from 0 to 2. In a specific embodiment, n' is 0 and n'' is 0. In a particular embodiment, n' is 1 and n'' is 0. In another embodiment, n' is 2 and n'' is 0. In yet another embodiment, n' is 3 and n'' is 0. In yet another embodiment, n' is 4 and n'' is 0. In yet another embodiment, n' is 5 and n'' is 0. In yet another embodiment, n' is 6 and n'' is 0. In a particular embodiment, n' is 0 and n'' is 1. In a particular embodiment, n' is 1 and n'' is 1. In another embodiment, n' is 2 and n'' is 1. In yet another embodiment, n' is 3 and n'' is 1. In yet another embodiment, n' is 4 and n'' is 1. In yet another embodiment, n' is 5 and n'' is 1. In yet another embodiment, n' is 6 and n'' is 1. In a particular embodiment, n' is 0 and n'' is 2. In a particular embodiment, n' is 1 and n'' is 2. In another embodiment, n' is 2 and n'' is 2. In yet another embodiment, n' is 3 and n'' is 2. In yet another embodiment, n' is 4 and n'' is 2. In yet another embodiment, n' is 5 and n'' is 2. In yet another embodiment, n' is 6 and n'' is 2. In a particular embodiment, n' is 0 and n'' is 3. In a particular embodiment, n' is 1 and n'' is 3. In another embodiment, n' is 2 and n'' is 3. In yet another embodiment, n' is 3 and n'' is 3. In yet another embodiment, n' is 4 and n'' is 3. In yet another embodiment, n' is 5 and n'' is 3. In yet another embodiment, n' is 6 and n'' is 3. In a specific embodiment, n' is 0 and n'' is 4. In a specific embodiment, n' is 1 and n'' is 4. In another embodiment, n' is 2 and n'' is 4. In yet another embodiment, n' is 3 and n'' is 4. In yet another embodiment, n' is 4 and n'' is 4. In yet another embodiment, n' is 5 and n'' is 4. In yet another embodiment, n' is 6 and n'' is 4. In a specific embodiment, n' is 0 and n'' is 5. In a specific embodiment, n' is 1 and n'' is 5. In another embodiment, n' is 2 and n'' is 5. In yet another embodiment, n' is 3 and n'' is 5. In yet another embodiment, n' is 4 and n'' is 5. In yet another embodiment, n' is 5 and n" is 5. In yet another embodiment, n' is 6 and n" is 5. In a particular embodiment, n' is 0 and n" is 6. In a particular embodiment, n' is 1 and n" is 6. In another embodiment, n' is 2 and n" is 6. In yet another embodiment, n' is 3 and n" is 6. In yet another embodiment, n' is 4 and n" is 6. In yet another embodiment, n' is 5 and n" is 6. In yet another embodiment, n' is 6 and n" is 6.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(XII)，其中X為CH 2(CH 2) n '，其中n'係選自0至10，且其中X'為CH 2O(CH 2) n ''CH 2，其中n''係選自0至10。在一實施例中，n'係選自0至5且n''係選自0至10。在一實施例中，n'係選自0至5且n''係選自0至5。在一實施例中，n'係選自0至3且n''係選自0至3。在一實施例中，n'係選自0至2且n''係選自0至2。在一特定實施例中，n'為0且n''為0。在一特定實施例中，n'為1且n''為0。在另一實施例中，n'為2且n''為0。在又一實施例中，n'為3且n''為0。在又另一實施例中，n'為4且n''為0。在又另一實施例中，n'為5且n''為0。在又另一實施例中，n'為6且n''為0。在一特定實施例中，n'為0且n''為1。在一特定實施例中，n'為1且n''為1。在另一實施例中，n'為2且n''為1。在又一實施例中，n'為3且n''為1。在又另一實施例中，n'為4且n''為1。在又另一實施例中，n'為5且n''為1。在又另一實施例中，n'為6且n''為1。在一特定實施例中，n'為0且n''為2。在一特定實施例中，n'為1且n''為2。在另一實施例中，n'為2且n''為2。在又一實施例中，n'為3且n''為2。在又另一實施例中，n'為4且n''為2。在又另一實施例中，n'為5且n''為2。在又另一實施例中，n'為6且n''為2。在一特定實施例中，n'為0且n''為3。在一特定實施例中，n'為1且n''為3。在另一實施例中，n'為2且n''為3。在又一實施例中，n'為3且n''為3。在又另一實施例中，n'為4且n''為3。在又另一實施例中，n'為5且n''為3。在又另一實施例中，n'為6且n''為3。在一特定實施例中，n'為0且n''為4。在一特定實施例中，n'為1且n''為4。在另一實施例中，n'為2且n''為4。在又一實施例中，n'為3且n''為4。在又另一實施例中，n'為4且n''為4。在又另一實施例中，n'為5且n''為4。在又另一實施例中，n'為6且n''為4。在一特定實施例中，n'為0且n''為5。在一特定實施例中，n'為1且n''為5。在另一實施例中，n'為2且n''為5。在又一實施例中，n'為3且n''為5。在又另一實施例中，n'為4且n''為5。在又另一實施例中，n'為5且n''為5。在又另一實施例中，n'為6且n''為5。在一特定實施例中，n'為0且n''為6。在一特定實施例中，n'為1且n''為6。在另一實施例中，n'為2且n''為6。在又一實施例中，n'為3且n''為6。在又另一實施例中，n'為4且n''為6。在又另一實施例中，n'為5且n''為6。在又另一實施例中，n'為6且n''為6。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (XII), wherein X is CH 2 (CH 2 ) n ' , wherein n' is selected from 0 to 10, and wherein X' is CH 2 O (CH 2 ) n '' CH 2 , wherein n'' is selected from 0 to 5 and n'' is selected from 0 to 10. In one embodiment, n' is selected from 0 to 5 and n'' is selected from 0 to 5. In one embodiment, n' is selected from 0 to 3 and n'' is selected from 0 to 3. In one embodiment, n' is selected from 0 to 2 and n'' is selected from 0 to 2. In a specific embodiment, n' is 0 and n'' is 0. In a particular embodiment, n' is 1 and n'' is 0. In another embodiment, n' is 2 and n'' is 0. In yet another embodiment, n' is 3 and n'' is 0. In yet another embodiment, n' is 4 and n'' is 0. In yet another embodiment, n' is 5 and n'' is 0. In yet another embodiment, n' is 6 and n'' is 0. In a particular embodiment, n' is 0 and n'' is 1. In a particular embodiment, n' is 1 and n'' is 1. In another embodiment, n' is 2 and n'' is 1. In yet another embodiment, n' is 3 and n'' is 1. In yet another embodiment, n' is 4 and n'' is 1. In yet another embodiment, n' is 5 and n'' is 1. In yet another embodiment, n' is 6 and n'' is 1. In a particular embodiment, n' is 0 and n'' is 2. In a particular embodiment, n' is 1 and n'' is 2. In another embodiment, n' is 2 and n'' is 2. In yet another embodiment, n' is 3 and n'' is 2. In yet another embodiment, n' is 4 and n'' is 2. In yet another embodiment, n' is 5 and n'' is 2. In yet another embodiment, n' is 6 and n'' is 2. In a particular embodiment, n' is 0 and n'' is 3. In a particular embodiment, n' is 1 and n'' is 3. In another embodiment, n' is 2 and n'' is 3. In yet another embodiment, n' is 3 and n'' is 3. In yet another embodiment, n' is 4 and n'' is 3. In yet another embodiment, n' is 5 and n'' is 3. In yet another embodiment, n' is 6 and n'' is 3. In a specific embodiment, n' is 0 and n'' is 4. In a specific embodiment, n' is 1 and n'' is 4. In another embodiment, n' is 2 and n'' is 4. In yet another embodiment, n' is 3 and n'' is 4. In yet another embodiment, n' is 4 and n'' is 4. In yet another embodiment, n' is 5 and n'' is 4. In yet another embodiment, n' is 6 and n'' is 4. In a specific embodiment, n' is 0 and n'' is 5. In a specific embodiment, n' is 1 and n'' is 5. In another embodiment, n' is 2 and n'' is 5. In yet another embodiment, n' is 3 and n'' is 5. In yet another embodiment, n' is 4 and n'' is 5. In yet another embodiment, n' is 5 and n" is 5. In yet another embodiment, n' is 6 and n" is 5. In a particular embodiment, n' is 0 and n" is 6. In a particular embodiment, n' is 1 and n" is 6. In another embodiment, n' is 2 and n" is 6. In yet another embodiment, n' is 3 and n" is 6. In yet another embodiment, n' is 4 and n" is 6. In yet another embodiment, n' is 5 and n" is 6. In yet another embodiment, n' is 6 and n" is 6.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(XII)，其中X為CH 2(CH 2) n '，其中n'係選自0至10，且其中X'為CH 2O(CH 2CH 2O) m '(CH 2) n ''CH 2，其中n''係選自0至10且m'係選自0至4。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (XII), wherein X is CH 2 (CH 2 ) n ′ , wherein n′ is selected from 0 to 10, and wherein X′ is CH 2 O(CH 2 CH 2 O) m ′ (CH 2 ) n ′′ CH 2 , wherein n′ is selected from 0 to 10 and m′ is selected from 0 to 4.

在一實施例中，n'係選自0至5，m'係選自0至4且n''係選自0至10。在一實施例中，n'係選自0至5，m'係選自0至4且n''係選自0至5。在一實施例中，n'係選自0至3，m'係選自0至2且n''係選自0至3。在一實施例中，n'係選自0至2，m'係選自0至2且n''係選自0至1。在一實施例中，n'係選自0至1，m'係選自0至1且n''係選自0至1。In one embodiment, n' is selected from 0 to 5, m' is selected from 0 to 4 and n'' is selected from 0 to 10. In one embodiment, n' is selected from 0 to 5, m' is selected from 0 to 4 and n'' is selected from 0 to 5. In one embodiment, n' is selected from 0 to 3, m' is selected from 0 to 2 and n'' is selected from 0 to 3. In one embodiment, n' is selected from 0 to 2, m' is selected from 0 to 2 and n'' is selected from 0 to 1. In one embodiment, n' is selected from 0 to 1, m' is selected from 0 to 1 and n'' is selected from 0 to 1.

在一特定實施例中，n'為0，m'為0且n''為0。在另一實施例中，n'為0，m'為1且n''為0。在另一實施例中，n'為0，m'為2且n''為0。在另一實施例中，n'為1，m'為3且n''為0。In a particular embodiment, n' is 0, m' is 0 and n" is 0. In another embodiment, n' is 0, m' is 1 and n" is 0. In another embodiment, n' is 0, m' is 2 and n" is 0. In another embodiment, n' is 1, m' is 3 and n" is 0.

在一特定實施例中，n'為1，m'為0且n''為0。在另一實施例中，n'為1，m'為1且n''為0。在另一實施例中，n'為1，m'為2且n''為0。在另一實施例中，n'為1，m'為3且n''為0。In a particular embodiment, n' is 1, m' is 0 and n'' is 0. In another embodiment, n' is 1, m' is 1 and n'' is 0. In another embodiment, n' is 1, m' is 2 and n'' is 0. In another embodiment, n' is 1, m' is 3 and n'' is 0.

在另一實施例中，n'為2，m'為0且n''為0。在另一實施例中，n'為2，m'為1且n''為0。在另一實施例中，n'為2，m'為2且n''為0。在另一實施例中，n'為2，m'為3且n''為0。In another embodiment, n' is 2, m' is 0 and n'' is 0. In another embodiment, n' is 2, m' is 1 and n'' is 0. In another embodiment, n' is 2, m' is 2 and n'' is 0. In another embodiment, n' is 2, m' is 3 and n'' is 0.

在又一實施例中，n'為3，m'為0且n''為0。在又一實施例中，n'為3，m'為1且n''為0。在又一實施例中，n'為3，m'為2且n''為0。在又一實施例中，n'為3，m'為3且n''為0。In yet another embodiment, n' is 3, m' is 0 and n'' is 0. In yet another embodiment, n' is 3, m' is 1 and n'' is 0. In yet another embodiment, n' is 3, m' is 2 and n'' is 0. In yet another embodiment, n' is 3, m' is 3 and n'' is 0.

在又另一實施例中，n'為4，m'為0且n''為0。在又另一實施例中，n'為4，m'為1且n''為0。在又另一實施例中，n'為4，m'為2且n''為0。在又另一實施例中，n'為4，m'為3且n''為0。In yet another embodiment, n' is 4, m' is 0 and n'' is 0. In yet another embodiment, n' is 4, m' is 1 and n'' is 0. In yet another embodiment, n' is 4, m' is 2 and n'' is 0. In yet another embodiment, n' is 4, m' is 3 and n'' is 0.

在又另一實施例中，n'為5，m'為0且n''為0。在又另一實施例中，n'為5，m'為1且n''為0。在又另一實施例中，n'為5，m'為2且n''為0。在又另一實施例中，n'為5，m'為3且n''為0。In yet another embodiment, n' is 5, m' is 0 and n'' is 0. In yet another embodiment, n' is 5, m' is 1 and n'' is 0. In yet another embodiment, n' is 5, m' is 2 and n'' is 0. In yet another embodiment, n' is 5, m' is 3 and n'' is 0.

在一特定實施例中，n'為0，m'為0且n''為1。在一特定實施例中，n'為0，m'為1且n''為1。在一特定實施例中，n'為0，m'為2且n''為1。在一特定實施例中，n'為0，m'為3且n''為1。In a specific embodiment, n' is 0, m' is 0 and n'' is 1. In a specific embodiment, n' is 0, m' is 1 and n'' is 1. In a specific embodiment, n' is 0, m' is 2 and n'' is 1. In a specific embodiment, n' is 0, m' is 3 and n'' is 1.

在一特定實施例中，n'為1，m'為0且n''為1。在一特定實施例中，n'為1，m'為1且n''為1。在一特定實施例中，n'為1，m'為2且n''為1。在一特定實施例中，n'為1，m'為3且n''為1。In a specific embodiment, n' is 1, m' is 0 and n'' is 1. In a specific embodiment, n' is 1, m' is 1 and n'' is 1. In a specific embodiment, n' is 1, m' is 2 and n'' is 1. In a specific embodiment, n' is 1, m' is 3 and n'' is 1.

在另一實施例中，n'為2，m'為0且n''為1。在另一實施例中，n'為2，m'為1且n''為1。在另一實施例中，n'為2，m'為2且n''為1。在另一實施例中，n'為2，m'為3且n''為1。In another embodiment, n' is 2, m' is 0 and n'' is 1. In another embodiment, n' is 2, m' is 1 and n'' is 1. In another embodiment, n' is 2, m' is 2 and n'' is 1. In another embodiment, n' is 2, m' is 3 and n'' is 1.

在又一實施例中，n'為3，m'為0且n''為1。在又一實施例中，n'為3，m'為1且n''為1。在又一實施例中，n'為3，m'為2且n''為1。在又一實施例中，n'為3，m'為3且n''為1。In yet another embodiment, n' is 3, m' is 0 and n'' is 1. In yet another embodiment, n' is 3, m' is 1 and n'' is 1. In yet another embodiment, n' is 3, m' is 2 and n'' is 1. In yet another embodiment, n' is 3, m' is 3 and n'' is 1.

在又另一實施例中，n'為4，m'為0且n''為1。在又另一實施例中，n'為4，m'為1且n''為1。在又另一實施例中，n'為4，m'為2且n''為1。在又另一實施例中，n'為4，m'為3且n''為1。In yet another embodiment, n' is 4, m' is 0 and n" is 1. In yet another embodiment, n' is 4, m' is 1 and n" is 1. In yet another embodiment, n' is 4, m' is 2 and n" is 1. In yet another embodiment, n' is 4, m' is 3 and n" is 1.

在又另一實施例中，n'為5，m'為0且n''為1。在又另一實施例中，n'為5，m'為1且n''為1。在又另一實施例中，n'為5，m'為2且n''為1。在又另一實施例中，n'為5，m'為3且n''為1。In yet another embodiment, n' is 5, m' is 0 and n'' is 1. In yet another embodiment, n' is 5, m' is 1 and n'' is 1. In yet another embodiment, n' is 5, m' is 2 and n'' is 1. In yet another embodiment, n' is 5, m' is 3 and n'' is 1.

在一特定實施例中，n'為0，m'為0且n''為2。在一特定實施例中，n'為0，m'為1且n''為2。在一特定實施例中，n'為0，m'為2且n''為2。在一特定實施例中，n'為0，m'為3且n''為2。In a specific embodiment, n' is 0, m' is 0 and n'' is 2. In a specific embodiment, n' is 0, m' is 1 and n'' is 2. In a specific embodiment, n' is 0, m' is 2 and n'' is 2. In a specific embodiment, n' is 0, m' is 3 and n'' is 2.

在一特定實施例中，n'為1，m'為0且n''為2。在一特定實施例中，n'為1，m'為1且n''為2。在一特定實施例中，n'為1，m'為2且n''為2。在一特定實施例中，n'為1，m'為3且n''為2。In a specific embodiment, n' is 1, m' is 0 and n'' is 2. In a specific embodiment, n' is 1, m' is 1 and n'' is 2. In a specific embodiment, n' is 1, m' is 2 and n'' is 2. In a specific embodiment, n' is 1, m' is 3 and n'' is 2.

在另一實施例中，n'為2，m'為0且n''為2。在另一實施例中，n'為2，m'為1且n''為2。在另一實施例中，n'為2，m'為2且n''為2。在另一實施例中，n'為2，m'為3且n''為2。In another embodiment, n' is 2, m' is 0 and n'' is 2. In another embodiment, n' is 2, m' is 1 and n'' is 2. In another embodiment, n' is 2, m' is 2 and n'' is 2. In another embodiment, n' is 2, m' is 3 and n'' is 2.

在又一實施例中，n'為3，m'為0且n''為2。在又一實施例中，n'為3，m'為1且n''為2。在又一實施例中，n'為3，m'為2且n''為2。在又一實施例中，n'為3，m'為3且n''為2。In yet another embodiment, n' is 3, m' is 0 and n'' is 2. In yet another embodiment, n' is 3, m' is 1 and n'' is 2. In yet another embodiment, n' is 3, m' is 2 and n'' is 2. In yet another embodiment, n' is 3, m' is 3 and n'' is 2.

在又另一實施例中，n'為4，m'為0且n''為2。在又另一實施例中，n'為4，m'為1且n''為2。在又另一實施例中，n'為4，m'為2且n''為2。在又另一實施例中，n'為4，m'為3且n''為2。In yet another embodiment, n' is 4, m' is 0 and n" is 2. In yet another embodiment, n' is 4, m' is 1 and n" is 2. In yet another embodiment, n' is 4, m' is 2 and n" is 2. In yet another embodiment, n' is 4, m' is 3 and n" is 2.

在又另一實施例中，n'為5，m'為0且n''為2。在又另一實施例中，n'為5，m'為1且n''為2。在又另一實施例中，n'為5，m'為2且n''為2。在又另一實施例中，n'為5，m'為3且n''為2。In yet another embodiment, n' is 5, m' is 0 and n'' is 2. In yet another embodiment, n' is 5, m' is 1 and n'' is 2. In yet another embodiment, n' is 5, m' is 2 and n'' is 2. In yet another embodiment, n' is 5, m' is 3 and n'' is 2.

在一特定實施例中，n'為0，m'為0且n''為3。在一特定實施例中，n'為0，m'為1且n''為3。在一特定實施例中，n'為0，m'為2且n''為3。在一特定實施例中，n'為0，m'為3且n''為3。In a specific embodiment, n' is 0, m' is 0 and n'' is 3. In a specific embodiment, n' is 0, m' is 1 and n'' is 3. In a specific embodiment, n' is 0, m' is 2 and n'' is 3. In a specific embodiment, n' is 0, m' is 3 and n'' is 3.

在一特定實施例中，n'為1，m'為0且n''為3。在一特定實施例中，n'為1，m'為1且n''為3。在一特定實施例中，n'為1，m'為2且n''為3。在一特定實施例中，n'為1，m'為3且n''為3。In a specific embodiment, n' is 1, m' is 0 and n'' is 3. In a specific embodiment, n' is 1, m' is 1 and n'' is 3. In a specific embodiment, n' is 1, m' is 2 and n'' is 3. In a specific embodiment, n' is 1, m' is 3 and n'' is 3.

在另一實施例中，n'為2，m'為0且n''為3。在另一實施例中，n'為2，m'為1且n''為3。在另一實施例中，n'為2，m'為2且n''為3。在另一實施例中，n'為2，m'為3且n''為3。In another embodiment, n' is 2, m' is 0 and n'' is 3. In another embodiment, n' is 2, m' is 1 and n'' is 3. In another embodiment, n' is 2, m' is 2 and n'' is 3. In another embodiment, n' is 2, m' is 3 and n'' is 3.

在又一實施例中，n'為3，m'為0且n''為3。在又一實施例中，n'為3，m'為1且n''為3。在又一實施例中，n'為3，m'為2且n''為3。在又一實施例中，n'為3，m'為3且n''為3。In yet another embodiment, n' is 3, m' is 0 and n'' is 3. In yet another embodiment, n' is 3, m' is 1 and n'' is 3. In yet another embodiment, n' is 3, m' is 2 and n'' is 3. In yet another embodiment, n' is 3, m' is 3 and n'' is 3.

在又另一實施例中，n'為4，m'為0且n''為3。在又另一實施例中，n'為4，m'為1且n''為3。在又另一實施例中，n'為4，m'為2且n''為3。在又另一實施例中，n'為4，m'為3且n''為3。In yet another embodiment, n' is 4, m' is 0 and n'' is 3. In yet another embodiment, n' is 4, m' is 1 and n'' is 3. In yet another embodiment, n' is 4, m' is 2 and n'' is 3. In yet another embodiment, n' is 4, m' is 3 and n'' is 3.

在又另一實施例中，n'為5，m'為0且n''為3。在又另一實施例中，n'為5，m'為1且n''為3。在又另一實施例中，n'為5，m'為2且n''為3。在又另一實施例中，n'為5，m'為3且n''為3。In yet another embodiment, n' is 5, m' is 0 and n'' is 3. In yet another embodiment, n' is 5, m' is 1 and n'' is 3. In yet another embodiment, n' is 5, m' is 2 and n'' is 3. In yet another embodiment, n' is 5, m' is 3 and n'' is 3.

在一特定實施例中，n'為0，m'為0且n''為4。在一特定實施例中，n'為0，m'為1且n''為4。在一特定實施例中，n'為0，m'為2且n''為4。在一特定實施例中，n'為0，m'為3且n''為4。In a specific embodiment, n' is 0, m' is 0 and n'' is 4. In a specific embodiment, n' is 0, m' is 1 and n'' is 4. In a specific embodiment, n' is 0, m' is 2 and n'' is 4. In a specific embodiment, n' is 0, m' is 3 and n'' is 4.

在一特定實施例中，n'為1，m'為0且n''為4。在一特定實施例中，n'為1，m'為1且n''為4。在一特定實施例中，n'為1，m'為2且n''為4。在一特定實施例中，n'為1，m'為3且n''為4。In a specific embodiment, n' is 1, m' is 0 and n'' is 4. In a specific embodiment, n' is 1, m' is 1 and n'' is 4. In a specific embodiment, n' is 1, m' is 2 and n'' is 4. In a specific embodiment, n' is 1, m' is 3 and n'' is 4.

在另一實施例中，n'為2，m'為0且n''為4。在另一實施例中，n'為2，m'為1且n''為4。在另一實施例中，n'為2，m'為2且n''為4。在另一實施例中，n'為2，m'為3且n''為4。In another embodiment, n' is 2, m' is 0 and n'' is 4. In another embodiment, n' is 2, m' is 1 and n'' is 4. In another embodiment, n' is 2, m' is 2 and n'' is 4. In another embodiment, n' is 2, m' is 3 and n'' is 4.

在又一實施例中，n'為3，m'為0且n''為4。在又一實施例中，n'為3，m'為1且n''為4。在又一實施例中，n'為3，m'為2且n''為4。在又一實施例中，n'為3，m'為3且n''為4。In yet another embodiment, n' is 3, m' is 0 and n'' is 4. In yet another embodiment, n' is 3, m' is 1 and n'' is 4. In yet another embodiment, n' is 3, m' is 2 and n'' is 4. In yet another embodiment, n' is 3, m' is 3 and n'' is 4.

在又另一實施例中，n'為4，m'為0且n''為4。在又另一實施例中，n'為4，m'為1且n''為4。在又另一實施例中，n'為4，m'為2且n''為4。在又另一實施例中，n'為4，m'為3且n''為4。In yet another embodiment, n' is 4, m' is 0 and n'' is 4. In yet another embodiment, n' is 4, m' is 1 and n'' is 4. In yet another embodiment, n' is 4, m' is 2 and n'' is 4. In yet another embodiment, n' is 4, m' is 3 and n'' is 4.

在又另一實施例中，n'為5，m'為0且n''為4。在又另一實施例中，n'為5，m'為1且n''為4。在又另一實施例中，n'為5，m'為2且n''為4。在又另一實施例中，n'為5，m'為3且n''為4。In yet another embodiment, n' is 5, m' is 0 and n'' is 4. In yet another embodiment, n' is 5, m' is 1 and n'' is 4. In yet another embodiment, n' is 5, m' is 2 and n'' is 4. In yet another embodiment, n' is 5, m' is 3 and n'' is 4.

在一特定實施例中，n'為0，m'為0且n''為5。在一特定實施例中，n'為0，m'為1且n''為5。在一特定實施例中，n'為0，m'為2且n''為5。在一特定實施例中，n'為0，m'為3且n''為5。In a specific embodiment, n' is 0, m' is 0 and n'' is 5. In a specific embodiment, n' is 0, m' is 1 and n'' is 5. In a specific embodiment, n' is 0, m' is 2 and n'' is 5. In a specific embodiment, n' is 0, m' is 3 and n'' is 5.

在一特定實施例中，n'為1，m'為0且n''為5。在一特定實施例中，n'為1，m'為1且n''為5。在一特定實施例中，n'為1，m'為2且n''為5。在一特定實施例中，n'為1，m'為3且n''為5。In a specific embodiment, n' is 1, m' is 0 and n'' is 5. In a specific embodiment, n' is 1, m' is 1 and n'' is 5. In a specific embodiment, n' is 1, m' is 2 and n'' is 5. In a specific embodiment, n' is 1, m' is 3 and n'' is 5.

在另一實施例中，n'為2，m'為0且n''為5。在另一實施例中，n'為2，m'為1且n''為5。在另一實施例中，n'為2，m'為2且n''為5。在另一實施例中，n'為2，m'為3且n''為5。In another embodiment, n' is 2, m' is 0 and n'' is 5. In another embodiment, n' is 2, m' is 1 and n'' is 5. In another embodiment, n' is 2, m' is 2 and n'' is 5. In another embodiment, n' is 2, m' is 3 and n'' is 5.

在又一實施例中，n'為3，m'為0且n''為5。在又一實施例中，n'為3，m'為1且n''為5。在又一實施例中，n'為3，m'為2且n''為5。在又一實施例中，n'為3，m'為3且n''為5。In yet another embodiment, n' is 3, m' is 0 and n'' is 5. In yet another embodiment, n' is 3, m' is 1 and n'' is 5. In yet another embodiment, n' is 3, m' is 2 and n'' is 5. In yet another embodiment, n' is 3, m' is 3 and n'' is 5.

在又另一實施例中，n'為4，m'為0且n''為5。在又另一實施例中，n'為4，m'為1且n''為5。在又另一實施例中，n'為4，m'為2且n''為5。在又另一實施例中，n'為4，m'為3且n''為5。In yet another embodiment, n' is 4, m' is 0 and n'' is 5. In yet another embodiment, n' is 4, m' is 1 and n'' is 5. In yet another embodiment, n' is 4, m' is 2 and n'' is 5. In yet another embodiment, n' is 4, m' is 3 and n'' is 5.

在又另一實施例中，n'為5，m'為0且n''為5。在又另一實施例中，n'為5，m'為1且n''為5。在又另一實施例中，n'為5，m'為2且n''為5。在又另一實施例中，n'為5，m'為3且n''為5。In yet another embodiment, n' is 5, m' is 0 and n'' is 5. In yet another embodiment, n' is 5, m' is 1 and n'' is 5. In yet another embodiment, n' is 5, m' is 2 and n'' is 5. In yet another embodiment, n' is 5, m' is 3 and n'' is 5.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(XII)，其中X為(CH 2CH 2O) mCH 2CH 2，其中m係選自1至4，且其中X'為CH 2(CH 2) n ''，其中n''係選自0至10。在一實施例中，m係選自1至3且n''係選自0至10。在一實施例中，m係選自1至3且n''係選自0至5。在一實施例中，m係選自1至2且n''係選自0至3。在一實施例中，m係選自1至2且n''係選自0至2。在一特定實施例中，m為1且n''為0。在另一實施例中，m為2且n''為0。在又一實施例中，m為3且n''為0。在又另一實施例中，m為4且n''為0。在一特定實施例中，m為1且n''為1。在另一實施例中，m為2且n''為1。在又一實施例中，m為3且n''為1。在又另一實施例中，m為4且n''為1。在一特定實施例中，m為1且n''為2。在另一實施例中，m為2且n''為2。在又一實施例中，m為3且n''為2。在又另一實施例中，m為4且n''為2。在一特定實施例中，m為1且n''為3。在另一實施例中，m為2且n''為3。在又一實施例中，m為3且n''為3。在又另一實施例中，m為4且n''為3。在一特定實施例中，m為1且n''為4。在另一實施例中，m為2且n''為4。在又一實施例中，m為3且n''為4。在又另一實施例中，m為4且n''為4。在一特定實施例中，m為1且n''為5。在另一實施例中，m為2且n''為5。在又一實施例中，m為3且n''為5。在又另一實施例中，m為4且n''為5。在一特定實施例中，m為1且n''為6。在另一實施例中，m為2且n''為6。在又一實施例中，m為3且n''為6。在又另一實施例中，m為4且n''為6。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (XII), wherein X is (CH 2 CH 2 O) m CH 2 CH 2 , wherein m is selected from 1 to 4, and wherein X' is CH 2 (CH 2 ) n '' , wherein n '' is selected from 0 to 10. In one embodiment, m is selected from 1 to 3 and n '' is selected from 0 to 10. In one embodiment, m is selected from 1 to 3 and n '' is selected from 0 to 5. In one embodiment, m is selected from 1 to 2 and n '' is selected from 0 to 3. In one embodiment, m is selected from 1 to 2 and n '' is selected from 0 to 2. In a specific embodiment, m is 1 and n '' is 0. In another embodiment, m is 2 and n'' is 0. In yet another embodiment, m is 3 and n'' is 0. In yet another embodiment, m is 4 and n'' is 0. In a specific embodiment, m is 1 and n'' is 1. In another embodiment, m is 2 and n'' is 1. In yet another embodiment, m is 3 and n'' is 1. In yet another embodiment, m is 4 and n'' is 1. In a specific embodiment, m is 1 and n'' is 2. In another embodiment, m is 2 and n'' is 2. In yet another embodiment, m is 3 and n'' is 2. In yet another embodiment, m is 4 and n'' is 2. In a specific embodiment, m is 1 and n'' is 3. In another embodiment, m is 2 and n'' is 3. In yet another embodiment, m is 3 and n'' is 3. In yet another embodiment, m is 4 and n'' is 3. In a specific embodiment, m is 1 and n'' is 4. In another embodiment, m is 2 and n'' is 4. In yet another embodiment, m is 3 and n'' is 4. In yet another embodiment, m is 4 and n'' is 4. In a specific embodiment, m is 1 and n'' is 5. In another embodiment, m is 2 and n'' is 5. In yet another embodiment, m is 3 and n'' is 5. In yet another embodiment, m is 4 and n'' is 5. In a specific embodiment, m is 1 and n'' is 6. In another embodiment, m is 2 and n'' is 6. In yet another embodiment, m is 3 and n'' is 6. In yet another embodiment, m is 4 and n″ is 6.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(XII)，其中X為(CH 2CH 2O) mCH 2CH 2，其中m係選自1至4，且其中X'為CH 2O(CH 2) n ''CH 2，其中n''係選自0至10。在一實施例中，m係選自1至3且n''係選自0至10。在一實施例中，m係選自1至3且n''係選自0至5。在一實施例中，m係選自1至2且n''係選自0至3。在一實施例中，m係選自1至2且n''係選自0至2。在一特定實施例中，m為1且n''為0。在另一實施例中，m為2且n''為0。在又一實施例中，m為3且n''為0。在又另一實施例中，m為4且n''為0。在一特定實施例中，m為1且n''為1。在另一實施例中，m為2且n''為1。在又一實施例中，m為3且n''為1。在又另一實施例中，m為4且n''為1。在一特定實施例中，m為1且n''為2。在另一實施例中，m為2且n''為2。在又一實施例中，m為3且n''為2。在又另一實施例中，m為4且n''為2。在一特定實施例中，m為1且n''為3。在另一實施例中，m為2且n''為3。在又一實施例中，m為3且n''為3。在又另一實施例中，m為4且n''為3。在一特定實施例中，m為1且n''為4。在另一實施例中，m為2且n''為4。在又一實施例中，m為3且n''為4。在又另一實施例中，m為4且n''為4。在一特定實施例中，m為1且n''為5。在另一實施例中，m為2且n''為5。在又一實施例中，m為3且n''為5。在又另一實施例中，m為4且n''為5。在一特定實施例中，m為1且n''為6。在另一實施例中，m為2且n''為6。在又一實施例中，m為3且n''為6。在又另一實施例中，m為4且n''為6。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (XII), wherein X is (CH 2 CH 2 O) m CH 2 CH 2 , wherein m is selected from 1 to 4, and wherein X' is CH 2 O(CH 2 ) n '' CH 2 , wherein n '' is selected from 0 to 10. In one embodiment, m is selected from 1 to 3 and n '' is selected from 0 to 10. In one embodiment, m is selected from 1 to 3 and n '' is selected from 0 to 5. In one embodiment, m is selected from 1 to 2 and n '' is selected from 0 to 3. In one embodiment, m is selected from 1 to 2 and n '' is selected from 0 to 2. In a specific embodiment, m is 1 and n '' is 0. In another embodiment, m is 2 and n'' is 0. In yet another embodiment, m is 3 and n'' is 0. In yet another embodiment, m is 4 and n'' is 0. In a specific embodiment, m is 1 and n'' is 1. In another embodiment, m is 2 and n'' is 1. In yet another embodiment, m is 3 and n'' is 1. In yet another embodiment, m is 4 and n'' is 1. In a specific embodiment, m is 1 and n'' is 2. In another embodiment, m is 2 and n'' is 2. In yet another embodiment, m is 3 and n'' is 2. In yet another embodiment, m is 4 and n'' is 2. In a specific embodiment, m is 1 and n'' is 3. In another embodiment, m is 2 and n'' is 3. In yet another embodiment, m is 3 and n'' is 3. In yet another embodiment, m is 4 and n'' is 3. In a specific embodiment, m is 1 and n'' is 4. In another embodiment, m is 2 and n'' is 4. In yet another embodiment, m is 3 and n'' is 4. In yet another embodiment, m is 4 and n'' is 4. In a specific embodiment, m is 1 and n'' is 5. In another embodiment, m is 2 and n'' is 5. In yet another embodiment, m is 3 and n'' is 5. In yet another embodiment, m is 4 and n'' is 5. In a specific embodiment, m is 1 and n'' is 6. In another embodiment, m is 2 and n'' is 6. In yet another embodiment, m is 3 and n'' is 6. In yet another embodiment, m is 4 and n″ is 6.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(XII)，其中X為(CH 2CH 2O) mCH 2CH 2，其中m係選自1至4，且其中X'為CH 2O(CH 2CH 2O) m '(CH 2) n ''CH 2，其中n''係選自0至10且m'係選自0至4。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (XII), wherein X is (CH 2 CH 2 O) m CH 2 CH 2 , wherein m is selected from 1 to 4, and wherein X' is CH 2 O(CH 2 CH 2 O) m ' (CH 2 ) n '' CH 2 , wherein n'' is selected from 0 to 10 and m' is selected from 0 to 4.

在一實施例中，m係選自1至3，m'係選自0至4且n''係選自0至10。在一實施例中，m係選自1至2，m'係選自0至4且n''係選自0至5。在一實施例中，m係選自1至2，m'係選自0至2且n''係選自0至3。在一實施例中，m係選自1至2，m'係選自0至2且n''係選自0至1。In one embodiment, m is selected from 1 to 3, m' is selected from 0 to 4 and n'' is selected from 0 to 10. In one embodiment, m is selected from 1 to 2, m' is selected from 0 to 4 and n'' is selected from 0 to 5. In one embodiment, m is selected from 1 to 2, m' is selected from 0 to 2 and n'' is selected from 0 to 3. In one embodiment, m is selected from 1 to 2, m' is selected from 0 to 2 and n'' is selected from 0 to 1.

在一特定實施例中，m為1，m'為0且n''為0。在另一實施例中，m為1，m'為1且n''為0。在另一實施例中，m為1，m'為2且n''為0。在另一實施例中，m為1，m'為3且n''為0。In a particular embodiment, m is 1, m' is 0 and n'' is 0. In another embodiment, m is 1, m' is 1 and n'' is 0. In another embodiment, m is 1, m' is 2 and n'' is 0. In another embodiment, m is 1, m' is 3 and n'' is 0.

在另一實施例中，m為2，m'為0且n''為0。在另一實施例中，m為2，m'為1且n''為0。在另一實施例中，m為2，m'為2且n''為0。在另一實施例中，m為2，m'為3且n''為0。In another embodiment, m is 2, m' is 0 and n'' is 0. In another embodiment, m is 2, m' is 1 and n'' is 0. In another embodiment, m is 2, m' is 2 and n'' is 0. In another embodiment, m is 2, m' is 3 and n'' is 0.

在又一實施例中，m為3，m'為0且n''為0。在又一實施例中，m為3，m'為1且n''為0。在又一實施例中，m為3，m'為2且n''為0。在又一實施例中，m為3，m'為3且n''為0。In yet another embodiment, m is 3, m' is 0 and n'' is 0. In yet another embodiment, m is 3, m' is 1 and n'' is 0. In yet another embodiment, m is 3, m' is 2 and n'' is 0. In yet another embodiment, m is 3, m' is 3 and n'' is 0.

在又另一實施例中，m為4，m'為0且n''為0。在又另一實施例中，m為4，m'為1且n''為0。在又另一實施例中，m為4，m'為2且n''為0。在又另一實施例中，m為4，m'為3且n''為0。In yet another embodiment, m is 4, m' is 0 and n" is 0. In yet another embodiment, m is 4, m' is 1 and n" is 0. In yet another embodiment, m is 4, m' is 2 and n" is 0. In yet another embodiment, m is 4, m' is 3 and n" is 0.

在一特定實施例中，m為1，m'為0且n''為1。在一特定實施例中，m為1，m'為1且n''為1。在一特定實施例中，m為1，m'為2且n''為1。在一特定實施例中，m為1，m'為3且n''為1。In a specific embodiment, m is 1, m' is 0 and n'' is 1. In a specific embodiment, m is 1, m' is 1 and n'' is 1. In a specific embodiment, m is 1, m' is 2 and n'' is 1. In a specific embodiment, m is 1, m' is 3 and n'' is 1.

在另一實施例中，m為2，m'為0且n''為1。在另一實施例中，m為2，m'為1且n''為1。在另一實施例中，m為2，m'為2且n''為1。在另一實施例中，m為2，m'為3且n''為1。In another embodiment, m is 2, m' is 0 and n'' is 1. In another embodiment, m is 2, m' is 1 and n'' is 1. In another embodiment, m is 2, m' is 2 and n'' is 1. In another embodiment, m is 2, m' is 3 and n'' is 1.

在又一實施例中，m為3，m'為0且n''為1。在又一實施例中，m為3，m'為1且n''為1。在又一實施例中，m為3，m'為2且n''為1。在又一實施例中，m為3，m'為3且n''為1。In yet another embodiment, m is 3, m' is 0 and n'' is 1. In yet another embodiment, m is 3, m' is 1 and n'' is 1. In yet another embodiment, m is 3, m' is 2 and n'' is 1. In yet another embodiment, m is 3, m' is 3 and n'' is 1.

在又另一實施例中，m為4，m'為0且n''為1。在又另一實施例中，m為4，m'為1且n''為1。在又另一實施例中，m為4，m'為2且n''為1。在又另一實施例中，m為4，m'為3且n''為1。In yet another embodiment, m is 4, m' is 0 and n'' is 1. In yet another embodiment, m is 4, m' is 1 and n'' is 1. In yet another embodiment, m is 4, m' is 2 and n'' is 1. In yet another embodiment, m is 4, m' is 3 and n'' is 1.

在一特定實施例中，m為1，m'為0且n''為2。在一特定實施例中，m為1，m'為1且n''為2。在一特定實施例中，m為1，m'為2且n''為2。在一特定實施例中，m為1，m'為3且n''為2。In a specific embodiment, m is 1, m' is 0 and n'' is 2. In a specific embodiment, m is 1, m' is 1 and n'' is 2. In a specific embodiment, m is 1, m' is 2 and n'' is 2. In a specific embodiment, m is 1, m' is 3 and n'' is 2.

在另一實施例中，m為2，m'為0且n''為2。在另一實施例中，m為2，m'為1且n''為2。在另一實施例中，m為2，m'為2且n''為2。在另一實施例中，m為2，m'為3且n''為2。In another embodiment, m is 2, m' is 0 and n'' is 2. In another embodiment, m is 2, m' is 1 and n'' is 2. In another embodiment, m is 2, m' is 2 and n'' is 2. In another embodiment, m is 2, m' is 3 and n'' is 2.

在又一實施例中，m為3，m'為0且n''為2。在又一實施例中，m為3，m'為1且n''為2。在又一實施例中，m為3，m'為2且n''為2。在又一實施例中，m為3，m'為3且n''為2。In yet another embodiment, m is 3, m' is 0 and n'' is 2. In yet another embodiment, m is 3, m' is 1 and n'' is 2. In yet another embodiment, m is 3, m' is 2 and n'' is 2. In yet another embodiment, m is 3, m' is 3 and n'' is 2.

在又另一實施例中，m為4，m'為0且n''為2。在又另一實施例中，m為4，m'為1且n''為2。在又另一實施例中，m為4，m'為2且n''為2。在又另一實施例中，m為4，m'為3且n''為2。In yet another embodiment, m is 4, m' is 0 and n'' is 2. In yet another embodiment, m is 4, m' is 1 and n'' is 2. In yet another embodiment, m is 4, m' is 2 and n'' is 2. In yet another embodiment, m is 4, m' is 3 and n'' is 2.

在一特定實施例中，m為1，m'為0且n''為3。在一特定實施例中，m為1，m'為1且n''為3。在一特定實施例中，m為1，m'為2且n''為3。在一特定實施例中，m為1，m'為3且n''為3。In a specific embodiment, m is 1, m' is 0 and n'' is 3. In a specific embodiment, m is 1, m' is 1 and n'' is 3. In a specific embodiment, m is 1, m' is 2 and n'' is 3. In a specific embodiment, m is 1, m' is 3 and n'' is 3.

在另一實施例中，m為2，m'為0且n''為3。在另一實施例中，m為2，m'為1且n''為3。在另一實施例中，m為2，m'為2且n''為3。在另一實施例中，m為2，m'為3且n''為3。In another embodiment, m is 2, m' is 0 and n'' is 3. In another embodiment, m is 2, m' is 1 and n'' is 3. In another embodiment, m is 2, m' is 2 and n'' is 3. In another embodiment, m is 2, m' is 3 and n'' is 3.

在又一實施例中，m為3，m'為0且n''為3。在又一實施例中，m為3，m'為1且n''為3。在又一實施例中，m為3，m'為2且n''為3。在又一實施例中，m為3，m'為3且n''為3。In yet another embodiment, m is 3, m' is 0 and n'' is 3. In yet another embodiment, m is 3, m' is 1 and n'' is 3. In yet another embodiment, m is 3, m' is 2 and n'' is 3. In yet another embodiment, m is 3, m' is 3 and n'' is 3.

在又另一實施例中，m為4，m'為0且n''為3。在又另一實施例中，m為4，m'為1且n''為3。在又另一實施例中，m為4，m'為2且n''為3。在又另一實施例中，m為4，m'為3且n''為3。In yet another embodiment, m is 4, m' is 0 and n'' is 3. In yet another embodiment, m is 4, m' is 1 and n'' is 3. In yet another embodiment, m is 4, m' is 2 and n'' is 3. In yet another embodiment, m is 4, m' is 3 and n'' is 3.

在一特定實施例中，m為1，m'為0且n''為4。在一特定實施例中，m為1，m'為1且n''為4。在一特定實施例中，m為1，m'為2且n''為4。在一特定實施例中，m為1，m'為3且n''為4。In a specific embodiment, m is 1, m' is 0 and n'' is 4. In a specific embodiment, m is 1, m' is 1 and n'' is 4. In a specific embodiment, m is 1, m' is 2 and n'' is 4. In a specific embodiment, m is 1, m' is 3 and n'' is 4.

在另一實施例中，m為2，m'為0且n''為4。在另一實施例中，m為2，m'為1且n''為4。在另一實施例中，m為2，m'為2且n''為4。在另一實施例中，m為2，m'為3且n''為4。In another embodiment, m is 2, m' is 0 and n'' is 4. In another embodiment, m is 2, m' is 1 and n'' is 4. In another embodiment, m is 2, m' is 2 and n'' is 4. In another embodiment, m is 2, m' is 3 and n'' is 4.

在又一實施例中，m為3，m'為0且n''為4。在又一實施例中，m為3，m'為1且n''為4。在又一實施例中，m為3，m'為2且n''為4。在又一實施例中，m為3，m'為3且n''為4。In yet another embodiment, m is 3, m' is 0 and n'' is 4. In yet another embodiment, m is 3, m' is 1 and n'' is 4. In yet another embodiment, m is 3, m' is 2 and n'' is 4. In yet another embodiment, m is 3, m' is 3 and n'' is 4.

在又另一實施例中，m為4，m'為0且n''為4。在又另一實施例中，m為4，m'為1且n''為4。在又另一實施例中，m為4，m'為2且n''為4。在又另一實施例中，m為4，m'為3且n''為4。In yet another embodiment, m is 4, m' is 0 and n'' is 4. In yet another embodiment, m is 4, m' is 1 and n'' is 4. In yet another embodiment, m is 4, m' is 2 and n'' is 4. In yet another embodiment, m is 4, m' is 3 and n'' is 4.

在一特定實施例中，m為1，m'為0且n''為5。在一特定實施例中，m為1，m'為1且n''為5。在一特定實施例中，m為1，m'為2且n''為5。在一特定實施例中，m為1，m'為3且n''為5。In a specific embodiment, m is 1, m' is 0 and n'' is 5. In a specific embodiment, m is 1, m' is 1 and n'' is 5. In a specific embodiment, m is 1, m' is 2 and n'' is 5. In a specific embodiment, m is 1, m' is 3 and n'' is 5.

在另一實施例中，m為2，m'為0且n''為5。在另一實施例中，m為2，m'為1且n''為5。在另一實施例中，m為2，m'為2且n''為5。在另一實施例中，m為2，m'為3且n''為5。In another embodiment, m is 2, m' is 0 and n'' is 5. In another embodiment, m is 2, m' is 1 and n'' is 5. In another embodiment, m is 2, m' is 2 and n'' is 5. In another embodiment, m is 2, m' is 3 and n'' is 5.

在又一實施例中，m為3，m'為0且n''為5。在又一實施例中，m為3，m'為1且n''為5。在又一實施例中，m為3，m'為2且n''為5。在又一實施例中，m為3，m'為3且n''為5。In yet another embodiment, m is 3, m' is 0 and n'' is 5. In yet another embodiment, m is 3, m' is 1 and n'' is 5. In yet another embodiment, m is 3, m' is 2 and n'' is 5. In yet another embodiment, m is 3, m' is 3 and n'' is 5.

在又另一實施例中，m為4，m'為0且n''為5。在又另一實施例中，m為4，m'為1且n''為5。在又另一實施例中，m為4，m'為2且n''為5。在又另一實施例中，m為4，m'為3且n''為5。In yet another embodiment, m is 4, m' is 0 and n'' is 5. In yet another embodiment, m is 4, m' is 1 and n'' is 5. In yet another embodiment, m is 4, m' is 2 and n'' is 5. In yet another embodiment, m is 4, m' is 3 and n'' is 5.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(XII)，其中X為NHCO(CH 2) n '，其中n'係選自1至10，且其中X'為CH 2(CH 2) n ''，其中n''係選自0至10。在一實施例中，n'係選自1至5且n''係選自0至10。在一實施例中，n'係選自1至5且n''係選自0至5。在一實施例中，n'係選自1至3且n''係選自0至3。在一實施例中，n'係選自1至2且n''係選自0至2。在一特定實施例中，n'為1且n''為0。在另一實施例中，n'為2且n''為0。在又一實施例中，n'為3且n''為0。在又另一實施例中，n'為4且n''為0。在又另一實施例中，n'為5且n''為0。在又另一實施例中，n'為6且n''為0。在一特定實施例中，n'為1且n''為1。在另一實施例中，n'為2且n''為1。在又一實施例中，n'為3且n''為1。在又另一實施例中，n'為4且n''為1。在又另一實施例中，n'為5且n''為1。在又另一實施例中，n'為6且n''為1。在一特定實施例中，n'為1且n''為2。在另一實施例中，n'為2且n''為2。在又一實施例中，n'為3且n''為2。在又另一實施例中，n'為4且n''為2。在又另一實施例中，n'為5且n''為2。在又另一實施例中，n'為6且n''為2。在一特定實施例中，n'為1且n''為3。在另一實施例中，n'為2且n''為3。在又一實施例中，n'為3且n''為3。在又另一實施例中，n'為4且n''為3。在又另一實施例中，n'為5且n''為3。在又另一實施例中，n'為6且n''為3。在一特定實施例中，n'為1且n''為4。在另一實施例中，n'為2且n''為4。在又一實施例中，n'為3且n''為4。在又另一實施例中，n'為4且n''為4。在又另一實施例中，n'為5且n''為4。在又另一實施例中，n'為6且n''為4。在一特定實施例中，n'為1且n''為5。在另一實施例中，n'為2且n''為5。在又一實施例中，n'為3且n''為5。在又另一實施例中，n'為4且n''為5。在又另一實施例中，n'為5且n''為5。在又另一實施例中，n'為6且n''為5。在一特定實施例中，n'為1且n''為6。在另一實施例中，n'為2且n''為6。在又一實施例中，n'為3且n''為6。在又另一實施例中，n'為4且n''為6。在又另一實施例中，n'為5且n''為6。在又另一實施例中，n'為6且n''為6。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (XII), wherein X is NHCO(CH 2 ) n ′ , wherein n′ is selected from 1 to 10, and wherein X′ is CH 2 (CH 2 ) n ′′ , wherein n′ is selected from 0 to 10. In one embodiment, n′ is selected from 1 to 5 and n′′ is selected from 0 to 10. In one embodiment, n′ is selected from 1 to 5 and n′′ is selected from 0 to 5. In one embodiment, n′ is selected from 1 to 3 and n′′ is selected from 0 to 3. In one embodiment, n′ is selected from 1 to 2 and n′′ is selected from 0 to 2. In a specific embodiment, n′ is 1 and n′′ is 0. In another embodiment, n' is 2 and n'' is 0. In yet another embodiment, n' is 3 and n'' is 0. In yet another embodiment, n' is 4 and n'' is 0. In yet another embodiment, n' is 5 and n'' is 0. In yet another embodiment, n' is 6 and n'' is 0. In a specific embodiment, n' is 1 and n'' is 1. In another embodiment, n' is 2 and n'' is 1. In yet another embodiment, n' is 3 and n'' is 1. In yet another embodiment, n' is 4 and n'' is 1. In yet another embodiment, n' is 5 and n'' is 1. In yet another embodiment, n' is 6 and n'' is 1. In a specific embodiment, n' is 1 and n'' is 2. In another embodiment, n' is 2 and n'' is 2. In yet another embodiment, n' is 3 and n'' is 2. In yet another embodiment, n' is 4 and n'' is 2. In yet another embodiment, n' is 5 and n'' is 2. In yet another embodiment, n' is 6 and n'' is 2. In a specific embodiment, n' is 1 and n'' is 3. In another embodiment, n' is 2 and n'' is 3. In yet another embodiment, n' is 3 and n'' is 3. In yet another embodiment, n' is 4 and n'' is 3. In yet another embodiment, n' is 5 and n'' is 3. In yet another embodiment, n' is 6 and n'' is 3. In a specific embodiment, n' is 1 and n'' is 4. In another embodiment, n' is 2 and n'' is 4. In yet another embodiment, n' is 3 and n'' is 4. In yet another embodiment, n' is 4 and n'' is 4. In yet another embodiment, n' is 5 and n'' is 4. In yet another embodiment, n' is 6 and n'' is 4. In a specific embodiment, n' is 1 and n'' is 5. In another embodiment, n' is 2 and n'' is 5. In yet another embodiment, n' is 3 and n'' is 5. In yet another embodiment, n' is 4 and n'' is 5. In yet another embodiment, n' is 5 and n'' is 5. In yet another embodiment, n' is 6 and n'' is 5. In a specific embodiment, n' is 1 and n'' is 6. In another embodiment, n' is 2 and n'' is 6. In yet another embodiment, n' is 3 and n'' is 6. In yet another embodiment, n' is 4 and n'' is 6. In yet another embodiment, n' is 5 and n" is 6. In yet another embodiment, n' is 6 and n" is 6.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(VII)，其中X為NHCO(CH 2) n '，其中n'係選自1至10，且其中X'為CH 2O(CH 2) n ''CH 2，其中n''係選自0至10。在一實施例中，n'係選自1至5且n''係選自0至10。在一實施例中，n'係選自1至5且n''係選自0至5。在一實施例中，n'係選自1至3且n''係選自0至3。在一實施例中，n'係選自1至2且n''係選自0至2。在一特定實施例中，n'為1且n''為0。在另一實施例中，n'為2且n''為0。在又一實施例中，n'為3且n''為0。在又另一實施例中，n'為4且n''為0。在又另一實施例中，n'為5且n''為0。在又另一實施例中，n'為6且n''為0。在一特定實施例中，n'為1且n''為1。在另一實施例中，n'為2且n''為1。在又一實施例中，n'為3且n''為1。在又另一實施例中，n'為4且n''為1。在又另一實施例中，n'為5且n''為1。在又另一實施例中，n'為6且n''為1。在一特定實施例中，n'為1且n''為2。在另一實施例中，n'為2且n''為2。在又一實施例中，n'為3且n''為2。在又另一實施例中，n'為4且n''為2。在又另一實施例中，n'為5且n''為2。在又另一實施例中，n'為6且n''為2。在一特定實施例中，n'為1且n''為3。在另一實施例中，n'為2且n''為3。在又一實施例中，n'為3且n''為3。在又另一實施例中，n'為4且n''為3。在又另一實施例中，n'為5且n''為3。在又另一實施例中，n'為6且n''為3。在一特定實施例中，n'為1且n''為4。在另一實施例中，n'為2且n''為4。在又一實施例中，n'為3且n''為4。在又另一實施例中，n'為4且n''為4。在又另一實施例中，n'為5且n''為4。在又另一實施例中，n'為6且n''為4。在一特定實施例中，n'為1且n''為5。在另一實施例中，n'為2且n''為5。在又一實施例中，n'為3且n''為5。在又另一實施例中，n'為4且n''為5。在又另一實施例中，n'為5且n''為5。在又另一實施例中，n'為6且n''為5。在一特定實施例中，n'為1且n''為6。在另一實施例中，n'為2且n''為6。在又一實施例中，n'為3且n''為6。在又另一實施例中，n'為4且n''為6。在又另一實施例中，n'為5且n''為6。在又另一實施例中，n'為6且n''為6。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (VII), wherein X is NHCO(CH 2 ) n ′ , wherein n′ is selected from 1 to 10, and wherein X′ is CH 2 O(CH 2 ) n ′′ CH 2 , wherein n′ is selected from 1 to 5 and n′′ is selected from 0 to 10. In one embodiment, n′ is selected from 1 to 5 and n′′ is selected from 0 to 10. In one embodiment, n′ is selected from 1 to 5 and n′′ is selected from 0 to 5. In one embodiment, n′ is selected from 1 to 3 and n′′ is selected from 0 to 3. In one embodiment, n′ is selected from 1 to 2 and n′′ is selected from 0 to 2. In a specific embodiment, n′ is 1 and n′′ is 0. In another embodiment, n' is 2 and n'' is 0. In yet another embodiment, n' is 3 and n'' is 0. In yet another embodiment, n' is 4 and n'' is 0. In yet another embodiment, n' is 5 and n'' is 0. In yet another embodiment, n' is 6 and n'' is 0. In a specific embodiment, n' is 1 and n'' is 1. In another embodiment, n' is 2 and n'' is 1. In yet another embodiment, n' is 3 and n'' is 1. In yet another embodiment, n' is 4 and n'' is 1. In yet another embodiment, n' is 5 and n'' is 1. In yet another embodiment, n' is 6 and n'' is 1. In a specific embodiment, n' is 1 and n'' is 2. In another embodiment, n' is 2 and n'' is 2. In yet another embodiment, n' is 3 and n'' is 2. In yet another embodiment, n' is 4 and n'' is 2. In yet another embodiment, n' is 5 and n'' is 2. In yet another embodiment, n' is 6 and n'' is 2. In a specific embodiment, n' is 1 and n'' is 3. In another embodiment, n' is 2 and n'' is 3. In yet another embodiment, n' is 3 and n'' is 3. In yet another embodiment, n' is 4 and n'' is 3. In yet another embodiment, n' is 5 and n'' is 3. In yet another embodiment, n' is 6 and n'' is 3. In a specific embodiment, n' is 1 and n'' is 4. In another embodiment, n' is 2 and n'' is 4. In yet another embodiment, n' is 3 and n'' is 4. In yet another embodiment, n' is 4 and n'' is 4. In yet another embodiment, n' is 5 and n'' is 4. In yet another embodiment, n' is 6 and n'' is 4. In a specific embodiment, n' is 1 and n'' is 5. In another embodiment, n' is 2 and n'' is 5. In yet another embodiment, n' is 3 and n'' is 5. In yet another embodiment, n' is 4 and n'' is 5. In yet another embodiment, n' is 5 and n'' is 5. In yet another embodiment, n' is 6 and n'' is 5. In a specific embodiment, n' is 1 and n'' is 6. In another embodiment, n' is 2 and n'' is 6. In yet another embodiment, n' is 3 and n'' is 6. In yet another embodiment, n' is 4 and n'' is 6. In yet another embodiment, n' is 5 and n" is 6. In yet another embodiment, n' is 6 and n" is 6.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(XII)，其中X為NHCO(CH 2) n '，其中n'係選自1至10，且其中X'為CH 2O(CH 2CH 2O) m '(CH 2) n ''CH 2，其中n''係選自0至10且m'係選自0至4。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (XII), wherein X is NHCO(CH 2 ) n ′ , wherein n′ is selected from 1 to 10, and wherein X′ is CH 2 O(CH 2 CH 2 O) m ′ (CH 2 ) n ′′ CH 2 , wherein n′ is selected from 0 to 10 and m′ is selected from 0 to 4.

在一實施例中，n'係選自1至5，m'係選自0至4且n''係選自0至10。在一實施例中，n'係選自1至5，m'係選自0至4且n''係選自0至5。在一實施例中，n'係選自1至3，m'係選自0至2且n''係選自0至3。在一實施例中，n'係選自1至2，m'係選自0至2且n''係選自0至1。In one embodiment, n' is selected from 1 to 5, m' is selected from 0 to 4 and n'' is selected from 0 to 10. In one embodiment, n' is selected from 1 to 5, m' is selected from 0 to 4 and n'' is selected from 0 to 5. In one embodiment, n' is selected from 1 to 3, m' is selected from 0 to 2 and n'' is selected from 0 to 3. In one embodiment, n' is selected from 1 to 2, m' is selected from 0 to 2 and n'' is selected from 0 to 1.

在一特定實施例中，n'為1，m'為0且n''為0。在另一實施例中，n'為1，m'為1且n''為0。在另一實施例中，n'為1，m'為2且n''為0。在另一實施例中，n'為1，m'為3且n''為0。In a particular embodiment, n' is 1, m' is 0 and n'' is 0. In another embodiment, n' is 1, m' is 1 and n'' is 0. In another embodiment, n' is 1, m' is 2 and n'' is 0. In another embodiment, n' is 1, m' is 3 and n'' is 0.

在另一實施例中，n'為2，m'為0且n''為0。在另一實施例中，n'為2，m'為1且n''為0。在另一實施例中，n'為2，m'為2且n''為0。在另一實施例中，n'為2，m'為3且n''為0。In another embodiment, n' is 2, m' is 0 and n'' is 0. In another embodiment, n' is 2, m' is 1 and n'' is 0. In another embodiment, n' is 2, m' is 2 and n'' is 0. In another embodiment, n' is 2, m' is 3 and n'' is 0.

在又一實施例中，n'為3，m'為0且n''為0。在又一實施例中，n'為3，m'為1且n''為0。在又一實施例中，n'為3，m'為2且n''為0。在又一實施例中，n'為3，m'為3且n''為0。In yet another embodiment, n' is 3, m' is 0 and n'' is 0. In yet another embodiment, n' is 3, m' is 1 and n'' is 0. In yet another embodiment, n' is 3, m' is 2 and n'' is 0. In yet another embodiment, n' is 3, m' is 3 and n'' is 0.

在又另一實施例中，n'為4，m'為0且n''為0。在又另一實施例中，n'為4，m'為1且n''為0。在又另一實施例中，n'為4，m'為2且n''為0。在又另一實施例中，n'為4，m'為3且n''為0。In yet another embodiment, n' is 4, m' is 0 and n'' is 0. In yet another embodiment, n' is 4, m' is 1 and n'' is 0. In yet another embodiment, n' is 4, m' is 2 and n'' is 0. In yet another embodiment, n' is 4, m' is 3 and n'' is 0.

在又另一實施例中，n'為5，m'為0且n''為0。在又另一實施例中，n'為5，m'為1且n''為0。在又另一實施例中，n'為5，m'為2且n''為0。在又另一實施例中，n'為5，m'為3且n''為0。In yet another embodiment, n' is 5, m' is 0 and n'' is 0. In yet another embodiment, n' is 5, m' is 1 and n'' is 0. In yet another embodiment, n' is 5, m' is 2 and n'' is 0. In yet another embodiment, n' is 5, m' is 3 and n'' is 0.

在一特定實施例中，n'為1，m'為0且n''為1。在一特定實施例中，n'為1，m'為1且n''為1。在一特定實施例中，n'為1，m'為2且n''為1。在一特定實施例中，n'為1，m'為3且n''為1。In a specific embodiment, n' is 1, m' is 0 and n'' is 1. In a specific embodiment, n' is 1, m' is 1 and n'' is 1. In a specific embodiment, n' is 1, m' is 2 and n'' is 1. In a specific embodiment, n' is 1, m' is 3 and n'' is 1.

在另一實施例中，n'為2，m'為0且n''為1。在另一實施例中，n'為2，m'為1且n''為1。在另一實施例中，n'為2，m'為2且n''為1。在另一實施例中，n'為2，m'為3且n''為1。In another embodiment, n' is 2, m' is 0 and n'' is 1. In another embodiment, n' is 2, m' is 1 and n'' is 1. In another embodiment, n' is 2, m' is 2 and n'' is 1. In another embodiment, n' is 2, m' is 3 and n'' is 1.

在又一實施例中，n'為3，m'為0且n''為1。在又一實施例中，n'為3，m'為1且n''為1。在又一實施例中，n'為3，m'為2且n''為1。在又一實施例中，n'為3，m'為3且n''為1。In yet another embodiment, n' is 3, m' is 0 and n'' is 1. In yet another embodiment, n' is 3, m' is 1 and n'' is 1. In yet another embodiment, n' is 3, m' is 2 and n'' is 1. In yet another embodiment, n' is 3, m' is 3 and n'' is 1.

在又另一實施例中，n'為4，m'為0且n''為1。在又另一實施例中，n'為4，m'為1且n''為1。在又另一實施例中，n'為4，m'為2且n''為1。在又另一實施例中，n'為4，m'為3且n''為1。In yet another embodiment, n' is 4, m' is 0 and n" is 1. In yet another embodiment, n' is 4, m' is 1 and n" is 1. In yet another embodiment, n' is 4, m' is 2 and n" is 1. In yet another embodiment, n' is 4, m' is 3 and n" is 1.

在又另一實施例中，n'為5，m'為0且n''為1。在又另一實施例中，n'為5，m'為1且n''為1。在又另一實施例中，n'為5，m'為2且n''為1。在又另一實施例中，n'為5，m'為3且n''為1。In yet another embodiment, n' is 5, m' is 0 and n'' is 1. In yet another embodiment, n' is 5, m' is 1 and n'' is 1. In yet another embodiment, n' is 5, m' is 2 and n'' is 1. In yet another embodiment, n' is 5, m' is 3 and n'' is 1.

在一特定實施例中，n'為1，m'為0且n''為2。在一特定實施例中，n'為1，m'為1且n''為2。在一特定實施例中，n'為1，m'為2且n''為2。在一特定實施例中，n'為1，m'為3且n''為2。In a specific embodiment, n' is 1, m' is 0 and n'' is 2. In a specific embodiment, n' is 1, m' is 1 and n'' is 2. In a specific embodiment, n' is 1, m' is 2 and n'' is 2. In a specific embodiment, n' is 1, m' is 3 and n'' is 2.

在另一實施例中，n'為2，m'為0且n''為2。在另一實施例中，n'為2，m'為1且n''為2。在另一實施例中，n'為2，m'為2且n''為2。在另一實施例中，n'為2，m'為3且n''為2。In another embodiment, n' is 2, m' is 0 and n'' is 2. In another embodiment, n' is 2, m' is 1 and n'' is 2. In another embodiment, n' is 2, m' is 2 and n'' is 2. In another embodiment, n' is 2, m' is 3 and n'' is 2.

在又一實施例中，n'為3，m'為0且n''為2。在又一實施例中，n'為3，m'為1且n''為2。在又一實施例中，n'為3，m'為2且n''為2。在又一實施例中，n'為3，m'為3且n''為2。In yet another embodiment, n' is 3, m' is 0 and n'' is 2. In yet another embodiment, n' is 3, m' is 1 and n'' is 2. In yet another embodiment, n' is 3, m' is 2 and n'' is 2. In yet another embodiment, n' is 3, m' is 3 and n'' is 2.

在又另一實施例中，n'為4，m'為0且n''為2。在又另一實施例中，n'為4，m'為1且n''為2。在又另一實施例中，n'為4，m'為2且n''為2。在又另一實施例中，n'為4，m'為3且n''為2。In yet another embodiment, n' is 4, m' is 0 and n" is 2. In yet another embodiment, n' is 4, m' is 1 and n" is 2. In yet another embodiment, n' is 4, m' is 2 and n" is 2. In yet another embodiment, n' is 4, m' is 3 and n" is 2.

在又另一實施例中，n'為5，m'為0且n''為2。在又另一實施例中，n'為5，m'為1且n''為2。在又另一實施例中，n'為5，m'為2且n''為2。在又另一實施例中，n'為5，m'為3且n''為2。In yet another embodiment, n' is 5, m' is 0 and n'' is 2. In yet another embodiment, n' is 5, m' is 1 and n'' is 2. In yet another embodiment, n' is 5, m' is 2 and n'' is 2. In yet another embodiment, n' is 5, m' is 3 and n'' is 2.

在一特定實施例中，n'為1，m'為0且n''為3。在一特定實施例中，n'為1，m'為1且n''為3。在一特定實施例中，n'為1，m'為2且n''為3。在一特定實施例中，n'為1，m'為3且n''為3。In a specific embodiment, n' is 1, m' is 0 and n'' is 3. In a specific embodiment, n' is 1, m' is 1 and n'' is 3. In a specific embodiment, n' is 1, m' is 2 and n'' is 3. In a specific embodiment, n' is 1, m' is 3 and n'' is 3.

在另一實施例中，n'為2，m'為0且n''為3。在另一實施例中，n'為2，m'為1且n''為3。在另一實施例中，n'為2，m'為2且n''為3。在另一實施例中，n'為2，m'為3且n''為3。In another embodiment, n' is 2, m' is 0 and n'' is 3. In another embodiment, n' is 2, m' is 1 and n'' is 3. In another embodiment, n' is 2, m' is 2 and n'' is 3. In another embodiment, n' is 2, m' is 3 and n'' is 3.

在又一實施例中，n'為3，m'為0且n''為3。在又一實施例中，n'為3，m'為1且n''為3。在又一實施例中，n'為3，m'為2且n''為3。在又一實施例中，n'為3，m'為3且n''為3。In yet another embodiment, n' is 3, m' is 0 and n'' is 3. In yet another embodiment, n' is 3, m' is 1 and n'' is 3. In yet another embodiment, n' is 3, m' is 2 and n'' is 3. In yet another embodiment, n' is 3, m' is 3 and n'' is 3.

在又另一實施例中，n'為4，m'為0且n''為3。在又另一實施例中，n'為4，m'為1且n''為3。在又另一實施例中，n'為4，m'為2且n''為3。在又另一實施例中，n'為4，m'為3且n''為3。In yet another embodiment, n' is 4, m' is 0 and n'' is 3. In yet another embodiment, n' is 4, m' is 1 and n'' is 3. In yet another embodiment, n' is 4, m' is 2 and n'' is 3. In yet another embodiment, n' is 4, m' is 3 and n'' is 3.

在又另一實施例中，n'為5，m'為0且n''為3。在又另一實施例中，n'為5，m'為1且n''為3。在又另一實施例中，n'為5，m'為2且n''為3。在又另一實施例中，n'為5，m'為3且n''為3。In yet another embodiment, n' is 5, m' is 0 and n'' is 3. In yet another embodiment, n' is 5, m' is 1 and n'' is 3. In yet another embodiment, n' is 5, m' is 2 and n'' is 3. In yet another embodiment, n' is 5, m' is 3 and n'' is 3.

在一特定實施例中，n'為1，m'為0且n''為4。在一特定實施例中，n'為1，m'為1且n''為4。在一特定實施例中，n'為1，m'為2且n''為4。在一特定實施例中，n'為1，m'為3且n''為4。In a specific embodiment, n' is 1, m' is 0 and n'' is 4. In a specific embodiment, n' is 1, m' is 1 and n'' is 4. In a specific embodiment, n' is 1, m' is 2 and n'' is 4. In a specific embodiment, n' is 1, m' is 3 and n'' is 4.

在另一實施例中，n'為2，m'為0且n''為4。在另一實施例中，n'為2，m'為1且n''為4。在另一實施例中，n'為2，m'為2且n''為4。在另一實施例中，n'為2，m'為3且n''為4。In another embodiment, n' is 2, m' is 0 and n'' is 4. In another embodiment, n' is 2, m' is 1 and n'' is 4. In another embodiment, n' is 2, m' is 2 and n'' is 4. In another embodiment, n' is 2, m' is 3 and n'' is 4.

在又一實施例中，n'為3，m'為0且n''為4。在又一實施例中，n'為3，m'為1且n''為4。在又一實施例中，n'為3，m'為2且n''為4。在又一實施例中，n'為3，m'為3且n''為4。In yet another embodiment, n' is 3, m' is 0 and n'' is 4. In yet another embodiment, n' is 3, m' is 1 and n'' is 4. In yet another embodiment, n' is 3, m' is 2 and n'' is 4. In yet another embodiment, n' is 3, m' is 3 and n'' is 4.

在又另一實施例中，n'為4，m'為0且n''為4。在又另一實施例中，n'為4，m'為1且n''為4。在又另一實施例中，n'為4，m'為2且n''為4。在又另一實施例中，n'為4，m'為3且n''為4。In yet another embodiment, n' is 4, m' is 0 and n'' is 4. In yet another embodiment, n' is 4, m' is 1 and n'' is 4. In yet another embodiment, n' is 4, m' is 2 and n'' is 4. In yet another embodiment, n' is 4, m' is 3 and n'' is 4.

在又另一實施例中，n'為5，m'為0且n''為4。在又另一實施例中，n'為5，m'為1且n''為4。在又另一實施例中，n'為5，m'為2且n''為4。在又另一實施例中，n'為5，m'為3且n''為4。In yet another embodiment, n' is 5, m' is 0 and n'' is 4. In yet another embodiment, n' is 5, m' is 1 and n'' is 4. In yet another embodiment, n' is 5, m' is 2 and n'' is 4. In yet another embodiment, n' is 5, m' is 3 and n'' is 4.

在一特定實施例中，n'為1，m'為0且n''為5。在一特定實施例中，n'為1，m'為1且n''為5。在一特定實施例中，n'為1，m'為2且n''為5。在一特定實施例中，n'為1，m'為3且n''為5。In a specific embodiment, n' is 1, m' is 0 and n'' is 5. In a specific embodiment, n' is 1, m' is 1 and n'' is 5. In a specific embodiment, n' is 1, m' is 2 and n'' is 5. In a specific embodiment, n' is 1, m' is 3 and n'' is 5.

在另一實施例中，n'為2，m'為0且n''為5。在另一實施例中，n'為2，m'為1且n''為5。在另一實施例中，n'為2，m'為2且n''為5。在另一實施例中，n'為2，m'為3且n''為5。In another embodiment, n' is 2, m' is 0 and n'' is 5. In another embodiment, n' is 2, m' is 1 and n'' is 5. In another embodiment, n' is 2, m' is 2 and n'' is 5. In another embodiment, n' is 2, m' is 3 and n'' is 5.

在又一實施例中，n'為3，m'為0且n''為5。在又一實施例中，n'為3，m'為1且n''為5。在又一實施例中，n'為3，m'為2且n''為5。在又一實施例中，n'為3，m'為3且n''為5。In yet another embodiment, n' is 3, m' is 0 and n'' is 5. In yet another embodiment, n' is 3, m' is 1 and n'' is 5. In yet another embodiment, n' is 3, m' is 2 and n'' is 5. In yet another embodiment, n' is 3, m' is 3 and n'' is 5.

在又另一實施例中，n'為4，m'為0且n''為5。在又另一實施例中，n'為4，m'為1且n''為5。在又另一實施例中，n'為4，m'為2且n''為5。在又另一實施例中，n'為4，m'為3且n''為5。In yet another embodiment, n' is 4, m' is 0 and n'' is 5. In yet another embodiment, n' is 4, m' is 1 and n'' is 5. In yet another embodiment, n' is 4, m' is 2 and n'' is 5. In yet another embodiment, n' is 4, m' is 3 and n'' is 5.

在又另一實施例中，n'為5，m'為0且n''為5。在又另一實施例中，n'為5，m'為1且n''為5。在又另一實施例中，n'為5，m'為2且n''為5。在又另一實施例中，n'為5，m'為3且n''為5。In yet another embodiment, n' is 5, m' is 0 and n'' is 5. In yet another embodiment, n' is 5, m' is 1 and n'' is 5. In yet another embodiment, n' is 5, m' is 2 and n'' is 5. In yet another embodiment, n' is 5, m' is 3 and n'' is 5.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(XII)，其中X為NHCO(CH 2CH 2O) mCH 2CH 2，其中m係選自1至4，且其中X'為CH 2(CH 2) n ''，其中n''係選自0至10。在一實施例中，m係選自1至3且n''係選自0至10。在一實施例中，m係選自1至3且n''係選自0至5。在一實施例中，m係選自1至2且n''係選自0至3。在一實施例中，m係選自1至2且n''係選自0至2。在一特定實施例中，m為1且n''為0。在另一實施例中，m為2且n''為0。在又一實施例中，m為3且n''為0。在又另一實施例中，m為4且n''為0。在一特定實施例中，m為1且n''為1。在另一實施例中，m為2且n''為1。在又一實施例中，m為3且n''為1。在又另一實施例中，m為4且n''為1。在一特定實施例中，m為1且n''為2。在另一實施例中，m為2且n''為2。在又一實施例中，m為3且n''為2。在又另一實施例中，m為4且n''為2。在一特定實施例中，m為1且n''為3。在另一實施例中，m為2且n''為3。在又一實施例中，m為3且n''為3。在又另一實施例中，m為4且n''為3。在一特定實施例中，m為1且n''為4。在另一實施例中，m為2且n''為4。在又一實施例中，m為3且n''為4。在又另一實施例中，m為4且n''為4。在一特定實施例中，m為1且n''為5。在另一實施例中，m為2且n''為5。在又一實施例中，m為3且n''為5。在又另一實施例中，m為4且n''為5。在一特定實施例中，m為1且n''為6。在另一實施例中，m為2且n''為6。在又一實施例中，m為3且n''為6。在又另一實施例中，m為4且n''為6。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (XII), wherein X is NHCO(CH 2 CH 2 O) m CH 2 CH 2 , wherein m is selected from 1 to 4, and wherein X' is CH 2 (CH 2 ) n '' , wherein n '' is selected from 0 to 10. In one embodiment, m is selected from 1 to 3 and n '' is selected from 0 to 10. In one embodiment, m is selected from 1 to 3 and n '' is selected from 0 to 5. In one embodiment, m is selected from 1 to 2 and n '' is selected from 0 to 3. In one embodiment, m is selected from 1 to 2 and n '' is selected from 0 to 2. In a specific embodiment, m is 1 and n '' is 0. In another embodiment, m is 2 and n'' is 0. In yet another embodiment, m is 3 and n'' is 0. In yet another embodiment, m is 4 and n'' is 0. In a specific embodiment, m is 1 and n'' is 1. In another embodiment, m is 2 and n'' is 1. In yet another embodiment, m is 3 and n'' is 1. In yet another embodiment, m is 4 and n'' is 1. In a specific embodiment, m is 1 and n'' is 2. In another embodiment, m is 2 and n'' is 2. In yet another embodiment, m is 3 and n'' is 2. In yet another embodiment, m is 4 and n'' is 2. In a specific embodiment, m is 1 and n'' is 3. In another embodiment, m is 2 and n'' is 3. In yet another embodiment, m is 3 and n'' is 3. In yet another embodiment, m is 4 and n'' is 3. In a specific embodiment, m is 1 and n'' is 4. In another embodiment, m is 2 and n'' is 4. In yet another embodiment, m is 3 and n'' is 4. In yet another embodiment, m is 4 and n'' is 4. In a specific embodiment, m is 1 and n'' is 5. In another embodiment, m is 2 and n'' is 5. In yet another embodiment, m is 3 and n'' is 5. In yet another embodiment, m is 4 and n'' is 5. In a specific embodiment, m is 1 and n'' is 6. In another embodiment, m is 2 and n'' is 6. In yet another embodiment, m is 3 and n'' is 6. In yet another embodiment, m is 4 and n″ is 6.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(XII)，其中X為NHCO(CH 2CH 2O) mCH 2CH 2，其中m係選自1至4，且其中X'為CH 2O(CH 2) n ''CH 2，其中n''係選自0至10。在一實施例中，m係選自1至3且n''係選自0至10。在一實施例中，m係選自1至3且n''係選自0至5。在一實施例中，m係選自1至2且n''係選自0至3。在一實施例中，m係選自1至2且n''係選自0至2。在一特定實施例中，m為1且n''為0。在另一實施例中，m為2且n''為0。在又一實施例中，m為3且n''為0。在又另一實施例中，m為4且n''為0。在一特定實施例中，m為1且n''為1。在另一實施例中，m為2且n''為1。在又一實施例中，m為3且n''為1。在又另一實施例中，m為4且n''為1。在一特定實施例中，m為1且n''為2。在另一實施例中，m為2且n''為2。在又一實施例中，m為3且n''為2。在又另一實施例中，m為4且n''為2。在一特定實施例中，m為1且n''為3。在另一實施例中，m為2且n''為3。在又一實施例中，m為3且n''為3。在又另一實施例中，m為4且n''為3。在一特定實施例中，m為1且n''為4。在另一實施例中，m為2且n''為4。在又一實施例中，m為3且n''為4。在又另一實施例中，m為4且n''為4。在一特定實施例中，m為1且n''為5。在另一實施例中，m為2且n''為5。在又一實施例中，m為3且n''為5。在又另一實施例中，m為4且n''為5。在一特定實施例中，m為1且n''為6。在另一實施例中，m為2且n''為6。在又一實施例中，m為3且n''為6。在又另一實施例中，m為4且n''為6。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (XII), wherein X is NHCO(CH 2 CH 2 O) m CH 2 CH 2 , wherein m is selected from 1 to 4, and wherein X' is CH 2 O(CH 2 ) n '' CH 2 , wherein n '' is selected from 0 to 10. In one embodiment, m is selected from 1 to 3 and n '' is selected from 0 to 10. In one embodiment, m is selected from 1 to 3 and n '' is selected from 0 to 5. In one embodiment, m is selected from 1 to 2 and n '' is selected from 0 to 3. In one embodiment, m is selected from 1 to 2 and n '' is selected from 0 to 2. In a specific embodiment, m is 1 and n '' is 0. In another embodiment, m is 2 and n'' is 0. In yet another embodiment, m is 3 and n'' is 0. In yet another embodiment, m is 4 and n'' is 0. In a specific embodiment, m is 1 and n'' is 1. In another embodiment, m is 2 and n'' is 1. In yet another embodiment, m is 3 and n'' is 1. In yet another embodiment, m is 4 and n'' is 1. In a specific embodiment, m is 1 and n'' is 2. In another embodiment, m is 2 and n'' is 2. In yet another embodiment, m is 3 and n'' is 2. In yet another embodiment, m is 4 and n'' is 2. In a specific embodiment, m is 1 and n'' is 3. In another embodiment, m is 2 and n'' is 3. In yet another embodiment, m is 3 and n'' is 3. In yet another embodiment, m is 4 and n'' is 3. In a specific embodiment, m is 1 and n'' is 4. In another embodiment, m is 2 and n'' is 4. In yet another embodiment, m is 3 and n'' is 4. In yet another embodiment, m is 4 and n'' is 4. In a specific embodiment, m is 1 and n'' is 5. In another embodiment, m is 2 and n'' is 5. In yet another embodiment, m is 3 and n'' is 5. In yet another embodiment, m is 4 and n'' is 5. In a specific embodiment, m is 1 and n'' is 6. In another embodiment, m is 2 and n'' is 6. In yet another embodiment, m is 3 and n'' is 6. In yet another embodiment, m is 4 and n″ is 6.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(XII)，其中X為NHCO(CH 2CH 2O) mCH 2CH 2，其中m係選自1至4，且其中X'為CH 2O(CH 2CH 2O) m '(CH 2) n ''CH 2，其中n''係選自0至10且m'係選自0至4。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (XII), wherein X is NHCO(CH 2 CH 2 O) m CH 2 CH 2 , wherein m is selected from 1 to 4, and wherein X' is CH 2 O(CH 2 CH 2 O) m ' (CH 2 ) n '' CH 2 , wherein n'' is selected from 0 to 10 and m' is selected from 0 to 4.

在一實施例中，m係選自1至3，m'係選自0至4且n''係選自0至10。在一實施例中，m係選自1至2，m'係選自0至4且n''係選自0至5。在一實施例中，m係選自1至2，m'係選自0至2且n''係選自0至3。在一實施例中，m係選自1至2，m'係選自0至2且n''係選自0至1。In one embodiment, m is selected from 1 to 3, m' is selected from 0 to 4 and n'' is selected from 0 to 10. In one embodiment, m is selected from 1 to 2, m' is selected from 0 to 4 and n'' is selected from 0 to 5. In one embodiment, m is selected from 1 to 2, m' is selected from 0 to 2 and n'' is selected from 0 to 3. In one embodiment, m is selected from 1 to 2, m' is selected from 0 to 2 and n'' is selected from 0 to 1.

在一特定實施例中，m為1，m'為0且n''為0。在另一實施例中，m為1，m'為1且n''為0。在另一實施例中，m為1，m'為2且n''為0。在另一實施例中，m為1，m'為3且n''為0。In a particular embodiment, m is 1, m' is 0 and n'' is 0. In another embodiment, m is 1, m' is 1 and n'' is 0. In another embodiment, m is 1, m' is 2 and n'' is 0. In another embodiment, m is 1, m' is 3 and n'' is 0.

在另一實施例中，m為2，m'為0且n''為0。在另一實施例中，m為2，m'為1且n''為0。在另一實施例中，m為2，m'為2且n''為0。在另一實施例中，m為2，m'為3且n''為0。In another embodiment, m is 2, m' is 0 and n'' is 0. In another embodiment, m is 2, m' is 1 and n'' is 0. In another embodiment, m is 2, m' is 2 and n'' is 0. In another embodiment, m is 2, m' is 3 and n'' is 0.

在又一實施例中，m為3，m'為0且n''為0。在又一實施例中，m為3，m'為1且n''為0。在又一實施例中，m為3，m'為2且n''為0。在又一實施例中，m為3，m'為3且n''為0。In yet another embodiment, m is 3, m' is 0 and n'' is 0. In yet another embodiment, m is 3, m' is 1 and n'' is 0. In yet another embodiment, m is 3, m' is 2 and n'' is 0. In yet another embodiment, m is 3, m' is 3 and n'' is 0.

在又另一實施例中，m為4，m'為0且n''為0。在又另一實施例中，m為4，m'為1且n''為0。在又另一實施例中，m為4，m'為2且n''為0。在又另一實施例中，m為4，m'為3且n''為0。In yet another embodiment, m is 4, m' is 0 and n" is 0. In yet another embodiment, m is 4, m' is 1 and n" is 0. In yet another embodiment, m is 4, m' is 2 and n" is 0. In yet another embodiment, m is 4, m' is 3 and n" is 0.

在一特定實施例中，m為1，m'為0且n''為1。在一特定實施例中，m為1，m'為1且n''為1。在一特定實施例中，m為1，m'為2且n''為1。在一特定實施例中，m為1，m'為3且n''為1。In a specific embodiment, m is 1, m' is 0 and n'' is 1. In a specific embodiment, m is 1, m' is 1 and n'' is 1. In a specific embodiment, m is 1, m' is 2 and n'' is 1. In a specific embodiment, m is 1, m' is 3 and n'' is 1.

在另一實施例中，m為2，m'為0且n''為1。在另一實施例中，m為2，m'為1且n''為1。在另一實施例中，m為2，m'為2且n''為1。在另一實施例中，m為2，m'為3且n''為1。In another embodiment, m is 2, m' is 0 and n'' is 1. In another embodiment, m is 2, m' is 1 and n'' is 1. In another embodiment, m is 2, m' is 2 and n'' is 1. In another embodiment, m is 2, m' is 3 and n'' is 1.

在又一實施例中，m為3，m'為0且n''為1。在又一實施例中，m為3，m'為1且n''為1。在又一實施例中，m為3，m'為2且n''為1。在又一實施例中，m為3，m'為3且n''為1。In yet another embodiment, m is 3, m' is 0 and n'' is 1. In yet another embodiment, m is 3, m' is 1 and n'' is 1. In yet another embodiment, m is 3, m' is 2 and n'' is 1. In yet another embodiment, m is 3, m' is 3 and n'' is 1.

在又另一實施例中，m為4，m'為0且n''為1。在又另一實施例中，m為4，m'為1且n''為1。在又另一實施例中，m為4，m'為2且n''為1。在又另一實施例中，m為4，m'為3且n''為1。In yet another embodiment, m is 4, m' is 0 and n'' is 1. In yet another embodiment, m is 4, m' is 1 and n'' is 1. In yet another embodiment, m is 4, m' is 2 and n'' is 1. In yet another embodiment, m is 4, m' is 3 and n'' is 1.

在一特定實施例中，m為1，m'為0且n''為2。在一特定實施例中，m為1，m'為1且n''為2。在一特定實施例中，m為1，m'為2且n''為2。在一特定實施例中，m為1，m'為3且n''為2。In a specific embodiment, m is 1, m' is 0 and n'' is 2. In a specific embodiment, m is 1, m' is 1 and n'' is 2. In a specific embodiment, m is 1, m' is 2 and n'' is 2. In a specific embodiment, m is 1, m' is 3 and n'' is 2.

在另一實施例中，m為2，m'為0且n''為2。在另一實施例中，m為2，m'為1且n''為2。在另一實施例中，m為2，m'為2且n''為2。在另一實施例中，m為2，m'為3且n''為2。In another embodiment, m is 2, m' is 0 and n'' is 2. In another embodiment, m is 2, m' is 1 and n'' is 2. In another embodiment, m is 2, m' is 2 and n'' is 2. In another embodiment, m is 2, m' is 3 and n'' is 2.

在又一實施例中，m為3，m'為0且n''為2。在又一實施例中，m為3，m'為1且n''為2。在又一實施例中，m為3，m'為2且n''為2。在又一實施例中，m為3，m'為3且n''為2。In yet another embodiment, m is 3, m' is 0 and n'' is 2. In yet another embodiment, m is 3, m' is 1 and n'' is 2. In yet another embodiment, m is 3, m' is 2 and n'' is 2. In yet another embodiment, m is 3, m' is 3 and n'' is 2.

在又另一實施例中，m為4，m'為0且n''為2。在又另一實施例中，m為4，m'為1且n''為2。在又另一實施例中，m為4，m'為2且n''為2。在又另一實施例中，m為4，m'為3且n''為2。In yet another embodiment, m is 4, m' is 0 and n'' is 2. In yet another embodiment, m is 4, m' is 1 and n'' is 2. In yet another embodiment, m is 4, m' is 2 and n'' is 2. In yet another embodiment, m is 4, m' is 3 and n'' is 2.

在一特定實施例中，m為1，m'為0且n''為3。在一特定實施例中，m為1，m'為1且n''為3。在一特定實施例中，m為1，m'為2且n''為3。在一特定實施例中，m為1，m'為3且n''為3。In a specific embodiment, m is 1, m' is 0 and n'' is 3. In a specific embodiment, m is 1, m' is 1 and n'' is 3. In a specific embodiment, m is 1, m' is 2 and n'' is 3. In a specific embodiment, m is 1, m' is 3 and n'' is 3.

在另一實施例中，m為2，m'為0且n''為3。在另一實施例中，m為2，m'為1且n''為3。在另一實施例中，m為2，m'為2且n''為3。在另一實施例中，m為2，m'為3且n''為3。In another embodiment, m is 2, m' is 0 and n'' is 3. In another embodiment, m is 2, m' is 1 and n'' is 3. In another embodiment, m is 2, m' is 2 and n'' is 3. In another embodiment, m is 2, m' is 3 and n'' is 3.

在又一實施例中，m為3，m'為0且n''為3。在又一實施例中，m為3，m'為1且n''為3。在又一實施例中，m為3，m'為2且n''為3。在又一實施例中，m為3，m'為3且n''為3。In yet another embodiment, m is 3, m' is 0 and n'' is 3. In yet another embodiment, m is 3, m' is 1 and n'' is 3. In yet another embodiment, m is 3, m' is 2 and n'' is 3. In yet another embodiment, m is 3, m' is 3 and n'' is 3.

在又另一實施例中，m為4，m'為0且n''為3。在又另一實施例中，m為4，m'為1且n''為3。在又另一實施例中，m為4，m'為2且n''為3。在又另一實施例中，m為4，m'為3且n''為3。In yet another embodiment, m is 4, m' is 0 and n'' is 3. In yet another embodiment, m is 4, m' is 1 and n'' is 3. In yet another embodiment, m is 4, m' is 2 and n'' is 3. In yet another embodiment, m is 4, m' is 3 and n'' is 3.

在一特定實施例中，m為1，m'為0且n''為4。在一特定實施例中，m為1，m'為1且n''為4。在一特定實施例中，m為1，m'為2且n''為4。在一特定實施例中，m為1，m'為3且n''為4。In a specific embodiment, m is 1, m' is 0 and n'' is 4. In a specific embodiment, m is 1, m' is 1 and n'' is 4. In a specific embodiment, m is 1, m' is 2 and n'' is 4. In a specific embodiment, m is 1, m' is 3 and n'' is 4.

在另一實施例中，m為2，m'為0且n''為4。在另一實施例中，m為2，m'為1且n''為4。在另一實施例中，m為2，m'為2且n''為4。在另一實施例中，m為2，m'為3且n''為4。In another embodiment, m is 2, m' is 0 and n'' is 4. In another embodiment, m is 2, m' is 1 and n'' is 4. In another embodiment, m is 2, m' is 2 and n'' is 4. In another embodiment, m is 2, m' is 3 and n'' is 4.

在又一實施例中，m為3，m'為0且n''為4。在又一實施例中，m為3，m'為1且n''為4。在又一實施例中，m為3，m'為2且n''為4。在又一實施例中，m為3，m'為3且n''為4。In yet another embodiment, m is 3, m' is 0 and n'' is 4. In yet another embodiment, m is 3, m' is 1 and n'' is 4. In yet another embodiment, m is 3, m' is 2 and n'' is 4. In yet another embodiment, m is 3, m' is 3 and n'' is 4.

在又另一實施例中，m為4，m'為0且n''為4。在又另一實施例中，m為4，m'為1且n''為4。在又另一實施例中，m為4，m'為2且n''為4。在又另一實施例中，m為4，m'為3且n''為4。In yet another embodiment, m is 4, m' is 0 and n'' is 4. In yet another embodiment, m is 4, m' is 1 and n'' is 4. In yet another embodiment, m is 4, m' is 2 and n'' is 4. In yet another embodiment, m is 4, m' is 3 and n'' is 4.

在一特定實施例中，m為1，m'為0且n''為5。在一特定實施例中，m為1，m'為1且n''為5。在一特定實施例中，m為1，m'為2且n''為5。在一特定實施例中，m為1，m'為3且n''為5。In a specific embodiment, m is 1, m' is 0 and n'' is 5. In a specific embodiment, m is 1, m' is 1 and n'' is 5. In a specific embodiment, m is 1, m' is 2 and n'' is 5. In a specific embodiment, m is 1, m' is 3 and n'' is 5.

在另一實施例中，m為2，m'為0且n''為5。在另一實施例中，m為2，m'為1且n''為5。在另一實施例中，m為2，m'為2且n''為5。在另一實施例中，m為2，m'為3且n''為5。In another embodiment, m is 2, m' is 0 and n'' is 5. In another embodiment, m is 2, m' is 1 and n'' is 5. In another embodiment, m is 2, m' is 2 and n'' is 5. In another embodiment, m is 2, m' is 3 and n'' is 5.

在又一實施例中，m為3，m'為0且n''為5。在又一實施例中，m為3，m'為1且n''為5。在又一實施例中，m為3，m'為2且n''為5。在又一實施例中，m為3，m'為3且n''為5。In yet another embodiment, m is 3, m' is 0 and n'' is 5. In yet another embodiment, m is 3, m' is 1 and n'' is 5. In yet another embodiment, m is 3, m' is 2 and n'' is 5. In yet another embodiment, m is 3, m' is 3 and n'' is 5.

在又另一實施例中，m為4，m'為0且n''為5。在又另一實施例中，m為4，m'為1且n''為5。在又另一實施例中，m為4，m'為2且n''為5。在又另一實施例中，m為4，m'為3且n''為5。In yet another embodiment, m is 4, m' is 0 and n'' is 5. In yet another embodiment, m is 4, m' is 1 and n'' is 5. In yet another embodiment, m is 4, m' is 2 and n'' is 5. In yet another embodiment, m is 4, m' is 3 and n'' is 5.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(XII)，其中X為OCH 2(CH 2) n '，其中n'係選自1至10，且其中X'為CH 2(CH 2) n ''，其中n''係選自0至10。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (XII), wherein X is OCH2 ( CH2 ) n ' , wherein n' is selected from 1 to 10, and wherein X' is CH2 ( CH2 ) n '' , wherein n'' is selected from 0 to 10.

在一實施例中，n'係選自1至5且n''係選自0至10。在一實施例中，n'係選自1至5且n''係選自0至5。在一實施例中，n'係選自1至3且n''係選自0至3。在一實施例中，n'係選自1至2且n''係選自0至2。在一特定實施例中，n'為1且n''為0。在另一實施例中，n'為2且n''為0。在又一實施例中，n'為3且n''為0。在又另一實施例中，n'為4且n''為0。在又另一實施例中，n'為5且n''為0。在又另一實施例中，n'為6且n''為0。在一特定實施例中，n'為1且n''為1。在另一實施例中，n'為2且n''為1。在又一實施例中，n'為3且n''為1。在又另一實施例中，n'為4且n''為1。在又另一實施例中，n'為5且n''為1。在又另一實施例中，n'為6且n''為1。在一特定實施例中，n'為1且n''為2。在另一實施例中，n'為2且n''為2。在又一實施例中，n'為3且n''為2。在又另一實施例中，n'為4且n''為2。在又另一實施例中，n'為5且n''為2。在又另一實施例中，n'為6且n''為2。在一特定實施例中，n'為1且n''為3。在另一實施例中，n'為2且n''為3。在又一實施例中，n'為3且n''為3。在又另一實施例中，n'為4且n''為3。在又另一實施例中，n'為5且n''為3。在又另一實施例中，n'為6且n''為3。在一特定實施例中，n'為1且n''為4。在另一實施例中，n'為2且n''為4。在又一實施例中，n'為3且n''為4。在又另一實施例中，n'為4且n''為4。在又另一實施例中，n'為5且n''為4。在又另一實施例中，n'為6且n''為4。在一特定實施例中，n'為1且n''為5。在另一實施例中，n'為2且n''為5。在又一實施例中，n'為3且n''為5。在又另一實施例中，n'為4且n''為5。在又另一實施例中，n'為5且n''為5。在又另一實施例中，n'為6且n''為5。在一特定實施例中，n'為1且n''為6。在另一實施例中，n'為2且n''為6。在又一實施例中，n'為3且n''為6。在又另一實施例中，n'為4且n''為6。在又另一實施例中，n'為5且n''為6。在又另一實施例中，n'為6且n''為6。In one embodiment, n' is selected from 1 to 5 and n'' is selected from 0 to 10. In one embodiment, n' is selected from 1 to 5 and n'' is selected from 0 to 5. In one embodiment, n' is selected from 1 to 3 and n'' is selected from 0 to 3. In one embodiment, n' is selected from 1 to 2 and n'' is selected from 0 to 2. In a specific embodiment, n' is 1 and n'' is 0. In another embodiment, n' is 2 and n'' is 0. In yet another embodiment, n' is 3 and n'' is 0. In yet another embodiment, n' is 4 and n'' is 0. In yet another embodiment, n' is 5 and n'' is 0. In yet another embodiment, n' is 6 and n'' is 0. In a specific embodiment, n' is 1 and n'' is 1. In another embodiment, n' is 2 and n'' is 1. In yet another embodiment, n' is 3 and n'' is 1. In yet another embodiment, n' is 4 and n'' is 1. In yet another embodiment, n' is 5 and n'' is 1. In yet another embodiment, n' is 6 and n'' is 1. In a specific embodiment, n' is 1 and n'' is 2. In another embodiment, n' is 2 and n'' is 2. In yet another embodiment, n' is 3 and n'' is 2. In yet another embodiment, n' is 4 and n'' is 2. In yet another embodiment, n' is 5 and n'' is 2. In yet another embodiment, n' is 6 and n'' is 2. In a specific embodiment, n' is 1 and n'' is 3. In another embodiment, n' is 2 and n'' is 3. In yet another embodiment, n' is 3 and n'' is 3. In yet another embodiment, n' is 4 and n'' is 3. In yet another embodiment, n' is 5 and n'' is 3. In yet another embodiment, n' is 6 and n'' is 3. In a specific embodiment, n' is 1 and n'' is 4. In another embodiment, n' is 2 and n'' is 4. In yet another embodiment, n' is 3 and n'' is 4. In yet another embodiment, n' is 4 and n'' is 4. In yet another embodiment, n' is 5 and n'' is 4. In yet another embodiment, n' is 6 and n'' is 4. In a specific embodiment, n' is 1 and n'' is 5. In another embodiment, n' is 2 and n'' is 5. In yet another embodiment, n' is 3 and n'' is 5. In yet another embodiment, n' is 4 and n" is 5. In yet another embodiment, n' is 5 and n" is 5. In yet another embodiment, n' is 6 and n" is 5. In a particular embodiment, n' is 1 and n" is 6. In another embodiment, n' is 2 and n" is 6. In yet another embodiment, n' is 3 and n" is 6. In yet another embodiment, n' is 4 and n" is 6. In yet another embodiment, n' is 5 and n" is 6. In yet another embodiment, n' is 6 and n" is 6.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(XII)，其中X為OCH 2(CH 2) n '，其中n'係選自1至10，且其中X'為CH 2O(CH 2) n ''CH 2，其中n''係選自0至10。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (XII), wherein X is OCH2 ( CH2 ) n ' , wherein n' is selected from 1 to 10, and wherein X' is CH2O ( CH2 )n''CH2 , wherein n'' is selected from 0 to 10.

在一實施例中，n'係選自1至5且n''係選自0至10。在一實施例中，n'係選自1至5且n''係選自0至5。在一實施例中，n'係選自1至3且n''係選自0至3。在一實施例中，n'係選自1至2且n''係選自0至2。在一特定實施例中，n'為1且n''為0。在另一實施例中，n'為2且n''為0。在又一實施例中，n'為3且n''為0。在又另一實施例中，n'為4且n''為0。在又另一實施例中，n'為5且n''為0。在又另一實施例中，n'為6且n''為0。在一特定實施例中，n'為1且n''為1。在另一實施例中，n'為2且n''為1。在又一實施例中，n'為3且n''為1。在又另一實施例中，n'為4且n''為1。在又另一實施例中，n'為5且n''為1。在又另一實施例中，n'為6且n''為1。在一特定實施例中，n'為1且n''為2。在另一實施例中，n'為2且n''為2。在又一實施例中，n'為3且n''為2。在又另一實施例中，n'為4且n''為2。在又另一實施例中，n'為5且n''為2。在又另一實施例中，n'為6且n''為2。在一特定實施例中，n'為1且n''為3。在另一實施例中，n'為2且n''為3。在又一實施例中，n'為3且n''為3。在又另一實施例中，n'為4且n''為3。在又另一實施例中，n'為5且n''為3。在又另一實施例中，n'為6且n''為3。在一特定實施例中，n'為1且n''為4。在另一實施例中，n'為2且n''為4。在又一實施例中，n'為3且n''為4。在又另一實施例中，n'為4且n''為4。在又另一實施例中，n'為5且n''為4。在又另一實施例中，n'為6且n''為4。在一特定實施例中，n'為1且n''為5。在另一實施例中，n'為2且n''為5。在又一實施例中，n'為3且n''為5。在又另一實施例中，n'為4且n''為5。在又另一實施例中，n'為5且n''為5。在又另一實施例中，n'為6且n''為5。在一特定實施例中，n'為1且n''為6。在另一實施例中，n'為2且n''為6。在又一實施例中，n'為3且n''為6。在又另一實施例中，n'為4且n''為6。在又另一實施例中，n'為5且n''為6。在又另一實施例中，n'為6且n''為6。In one embodiment, n' is selected from 1 to 5 and n'' is selected from 0 to 10. In one embodiment, n' is selected from 1 to 5 and n'' is selected from 0 to 5. In one embodiment, n' is selected from 1 to 3 and n'' is selected from 0 to 3. In one embodiment, n' is selected from 1 to 2 and n'' is selected from 0 to 2. In a specific embodiment, n' is 1 and n'' is 0. In another embodiment, n' is 2 and n'' is 0. In yet another embodiment, n' is 3 and n'' is 0. In yet another embodiment, n' is 4 and n'' is 0. In yet another embodiment, n' is 5 and n'' is 0. In yet another embodiment, n' is 6 and n'' is 0. In a specific embodiment, n' is 1 and n'' is 1. In another embodiment, n' is 2 and n'' is 1. In yet another embodiment, n' is 3 and n'' is 1. In yet another embodiment, n' is 4 and n'' is 1. In yet another embodiment, n' is 5 and n'' is 1. In yet another embodiment, n' is 6 and n'' is 1. In a specific embodiment, n' is 1 and n'' is 2. In another embodiment, n' is 2 and n'' is 2. In yet another embodiment, n' is 3 and n'' is 2. In yet another embodiment, n' is 4 and n'' is 2. In yet another embodiment, n' is 5 and n'' is 2. In yet another embodiment, n' is 6 and n'' is 2. In a specific embodiment, n' is 1 and n'' is 3. In another embodiment, n' is 2 and n'' is 3. In yet another embodiment, n' is 3 and n'' is 3. In yet another embodiment, n' is 4 and n'' is 3. In yet another embodiment, n' is 5 and n'' is 3. In yet another embodiment, n' is 6 and n'' is 3. In a specific embodiment, n' is 1 and n'' is 4. In another embodiment, n' is 2 and n'' is 4. In yet another embodiment, n' is 3 and n'' is 4. In yet another embodiment, n' is 4 and n'' is 4. In yet another embodiment, n' is 5 and n'' is 4. In yet another embodiment, n' is 6 and n'' is 4. In a specific embodiment, n' is 1 and n'' is 5. In another embodiment, n' is 2 and n'' is 5. In yet another embodiment, n' is 3 and n'' is 5. In yet another embodiment, n' is 4 and n" is 5. In yet another embodiment, n' is 5 and n" is 5. In yet another embodiment, n' is 6 and n" is 5. In a particular embodiment, n' is 1 and n" is 6. In another embodiment, n' is 2 and n" is 6. In yet another embodiment, n' is 3 and n" is 6. In yet another embodiment, n' is 4 and n" is 6. In yet another embodiment, n' is 5 and n" is 6. In yet another embodiment, n' is 6 and n" is 6.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(XII)，其中X為OCH 2(CH 2) n '，其中n'係選自1至10，且其中X'為CH 2O(CH 2CH 2O) m '(CH 2) n ''CH 2，其中n''係選自0至10且m'係選自0至4。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (XII), wherein X is OCH2 ( CH2 ) n ' , wherein n' is selected from 1 to 10, and wherein X' is CH2O (CH2CH2O ) m ' ( CH2 )n''CH2 , wherein n'' is selected from 0 to 10 and m' is selected from 0 to 4.

在一實施例中，n'係選自1至5，m'係選自0至4且n''係選自0至10。在一實施例中，n'係選自1至5，m'係選自0至4且n''係選自0至5。在一實施例中，n'係選自1至3，m'係選自0至2且n''係選自0至3。在一實施例中，n'係選自1至2，m'係選自0至2且n''係選自0至1。In one embodiment, n' is selected from 1 to 5, m' is selected from 0 to 4 and n'' is selected from 0 to 10. In one embodiment, n' is selected from 1 to 5, m' is selected from 0 to 4 and n'' is selected from 0 to 5. In one embodiment, n' is selected from 1 to 3, m' is selected from 0 to 2 and n'' is selected from 0 to 3. In one embodiment, n' is selected from 1 to 2, m' is selected from 0 to 2 and n'' is selected from 0 to 1.

在一特定實施例中，n'為1，m'為0且n''為0。在另一實施例中，n'為1，m'為1且n''為0。在另一實施例中，n'為1，m'為2且n''為0。在另一實施例中，n'為1，m'為3且n''為0。In a particular embodiment, n' is 1, m' is 0 and n'' is 0. In another embodiment, n' is 1, m' is 1 and n'' is 0. In another embodiment, n' is 1, m' is 2 and n'' is 0. In another embodiment, n' is 1, m' is 3 and n'' is 0.

在另一實施例中，n'為2，m'為0且n''為0。在另一實施例中，n'為2，m'為1且n''為0。在另一實施例中，n'為2，m'為2且n''為0。在另一實施例中，n'為2，m'為3且n''為0。In another embodiment, n' is 2, m' is 0 and n'' is 0. In another embodiment, n' is 2, m' is 1 and n'' is 0. In another embodiment, n' is 2, m' is 2 and n'' is 0. In another embodiment, n' is 2, m' is 3 and n'' is 0.

在又一實施例中，n'為3，m'為0且n''為0。在又一實施例中，n'為3，m'為1且n''為0。在又一實施例中，n'為3，m'為2且n''為0。在又一實施例中，n'為3，m'為3且n''為0。In yet another embodiment, n' is 3, m' is 0 and n'' is 0. In yet another embodiment, n' is 3, m' is 1 and n'' is 0. In yet another embodiment, n' is 3, m' is 2 and n'' is 0. In yet another embodiment, n' is 3, m' is 3 and n'' is 0.

在又另一實施例中，n'為4，m'為0且n''為0。在又另一實施例中，n'為4，m'為1且n''為0。在又另一實施例中，n'為4，m'為2且n''為0。在又另一實施例中，n'為4，m'為3且n''為0。In yet another embodiment, n' is 4, m' is 0 and n'' is 0. In yet another embodiment, n' is 4, m' is 1 and n'' is 0. In yet another embodiment, n' is 4, m' is 2 and n'' is 0. In yet another embodiment, n' is 4, m' is 3 and n'' is 0.

在又另一實施例中，n'為5，m'為0且n''為0。在又另一實施例中，n'為5，m'為1且n''為0。在又另一實施例中，n'為5，m'為2且n''為0。在又另一實施例中，n'為5，m'為3且n''為0。In yet another embodiment, n' is 5, m' is 0 and n'' is 0. In yet another embodiment, n' is 5, m' is 1 and n'' is 0. In yet another embodiment, n' is 5, m' is 2 and n'' is 0. In yet another embodiment, n' is 5, m' is 3 and n'' is 0.

在一特定實施例中，n'為1，m'為0且n''為1。在一特定實施例中，n'為1，m'為1且n''為1。在一特定實施例中，n'為1，m'為2且n''為1。在一特定實施例中，n'為1，m'為3且n''為1。In a specific embodiment, n' is 1, m' is 0 and n'' is 1. In a specific embodiment, n' is 1, m' is 1 and n'' is 1. In a specific embodiment, n' is 1, m' is 2 and n'' is 1. In a specific embodiment, n' is 1, m' is 3 and n'' is 1.

在另一實施例中，n'為2，m'為0且n''為1。在另一實施例中，n'為2，m'為1且n''為1。在另一實施例中，n'為2，m'為2且n''為1。在另一實施例中，n'為2，m'為3且n''為1。In another embodiment, n' is 2, m' is 0 and n'' is 1. In another embodiment, n' is 2, m' is 1 and n'' is 1. In another embodiment, n' is 2, m' is 2 and n'' is 1. In another embodiment, n' is 2, m' is 3 and n'' is 1.

在又一實施例中，n'為3，m'為0且n''為1。在又一實施例中，n'為3，m'為1且n''為1。在又一實施例中，n'為3，m'為2且n''為1。在又一實施例中，n'為3，m'為3且n''為1。In yet another embodiment, n' is 3, m' is 0 and n'' is 1. In yet another embodiment, n' is 3, m' is 1 and n'' is 1. In yet another embodiment, n' is 3, m' is 2 and n'' is 1. In yet another embodiment, n' is 3, m' is 3 and n'' is 1.

在又另一實施例中，n'為4，m'為0且n''為1。在又另一實施例中，n'為4，m'為1且n''為1。在又另一實施例中，n'為4，m'為2且n''為1。在又另一實施例中，n'為4，m'為3且n''為1。In yet another embodiment, n' is 4, m' is 0 and n" is 1. In yet another embodiment, n' is 4, m' is 1 and n" is 1. In yet another embodiment, n' is 4, m' is 2 and n" is 1. In yet another embodiment, n' is 4, m' is 3 and n" is 1.

在又另一實施例中，n'為5，m'為0且n''為1。在又另一實施例中，n'為5，m'為1且n''為1。在又另一實施例中，n'為5，m'為2且n''為1。在又另一實施例中，n'為5，m'為3且n''為1。In yet another embodiment, n' is 5, m' is 0 and n'' is 1. In yet another embodiment, n' is 5, m' is 1 and n'' is 1. In yet another embodiment, n' is 5, m' is 2 and n'' is 1. In yet another embodiment, n' is 5, m' is 3 and n'' is 1.

在一特定實施例中，n'為1，m'為0且n''為2。在一特定實施例中，n'為1，m'為1且n''為2。在一特定實施例中，n'為1，m'為2且n''為2。在一特定實施例中，n'為1，m'為3且n''為2。In a specific embodiment, n' is 1, m' is 0 and n'' is 2. In a specific embodiment, n' is 1, m' is 1 and n'' is 2. In a specific embodiment, n' is 1, m' is 2 and n'' is 2. In a specific embodiment, n' is 1, m' is 3 and n'' is 2.

在另一實施例中，n'為2，m'為0且n''為2。在另一實施例中，n'為2，m'為1且n''為2。在另一實施例中，n'為2，m'為2且n''為2。在另一實施例中，n'為2，m'為3且n''為2。In another embodiment, n' is 2, m' is 0 and n'' is 2. In another embodiment, n' is 2, m' is 1 and n'' is 2. In another embodiment, n' is 2, m' is 2 and n'' is 2. In another embodiment, n' is 2, m' is 3 and n'' is 2.

在又一實施例中，n'為3，m'為0且n''為2。在又一實施例中，n'為3，m'為1且n''為2。在又一實施例中，n'為3，m'為2且n''為2。在又一實施例中，n'為3，m'為3且n''為2。In yet another embodiment, n' is 3, m' is 0 and n'' is 2. In yet another embodiment, n' is 3, m' is 1 and n'' is 2. In yet another embodiment, n' is 3, m' is 2 and n'' is 2. In yet another embodiment, n' is 3, m' is 3 and n'' is 2.

在又另一實施例中，n'為4，m'為0且n''為2。在又另一實施例中，n'為4，m'為1且n''為2。在又另一實施例中，n'為4，m'為2且n''為2。在又另一實施例中，n'為4，m'為3且n''為2。In yet another embodiment, n' is 4, m' is 0 and n" is 2. In yet another embodiment, n' is 4, m' is 1 and n" is 2. In yet another embodiment, n' is 4, m' is 2 and n" is 2. In yet another embodiment, n' is 4, m' is 3 and n" is 2.

在又另一實施例中，n'為5，m'為0且n''為2。在又另一實施例中，n'為5，m'為1且n''為2。在又另一實施例中，n'為5，m'為2且n''為2。在又另一實施例中，n'為5，m'為3且n''為2。In yet another embodiment, n' is 5, m' is 0 and n'' is 2. In yet another embodiment, n' is 5, m' is 1 and n'' is 2. In yet another embodiment, n' is 5, m' is 2 and n'' is 2. In yet another embodiment, n' is 5, m' is 3 and n'' is 2.

在一特定實施例中，n'為1，m'為0且n''為3。在一特定實施例中，n'為1，m'為1且n''為3。在一特定實施例中，n'為1，m'為2且n''為3。在一特定實施例中，n'為1，m'為3且n''為3。In a specific embodiment, n' is 1, m' is 0 and n'' is 3. In a specific embodiment, n' is 1, m' is 1 and n'' is 3. In a specific embodiment, n' is 1, m' is 2 and n'' is 3. In a specific embodiment, n' is 1, m' is 3 and n'' is 3.

在另一實施例中，n'為2，m'為0且n''為3。在另一實施例中，n'為2，m'為1且n''為3。在另一實施例中，n'為2，m'為2且n''為3。在另一實施例中，n'為2，m'為3且n''為3。In another embodiment, n' is 2, m' is 0 and n'' is 3. In another embodiment, n' is 2, m' is 1 and n'' is 3. In another embodiment, n' is 2, m' is 2 and n'' is 3. In another embodiment, n' is 2, m' is 3 and n'' is 3.

在又一實施例中，n'為3，m'為0且n''為3。在又一實施例中，n'為3，m'為1且n''為3。在又一實施例中，n'為3，m'為2且n''為3。在又一實施例中，n'為3，m'為3且n''為3。In yet another embodiment, n' is 3, m' is 0 and n'' is 3. In yet another embodiment, n' is 3, m' is 1 and n'' is 3. In yet another embodiment, n' is 3, m' is 2 and n'' is 3. In yet another embodiment, n' is 3, m' is 3 and n'' is 3.

在又另一實施例中，n'為4，m'為0且n''為3。在又另一實施例中，n'為4，m'為1且n''為3。在又另一實施例中，n'為4，m'為2且n''為3。在又另一實施例中，n'為4，m'為3且n''為3。In yet another embodiment, n' is 4, m' is 0 and n'' is 3. In yet another embodiment, n' is 4, m' is 1 and n'' is 3. In yet another embodiment, n' is 4, m' is 2 and n'' is 3. In yet another embodiment, n' is 4, m' is 3 and n'' is 3.

在又另一實施例中，n'為5，m'為0且n''為3。在又另一實施例中，n'為5，m'為1且n''為3。在又另一實施例中，n'為5，m'為2且n''為3。在又另一實施例中，n'為5，m'為3且n''為3。In yet another embodiment, n' is 5, m' is 0 and n'' is 3. In yet another embodiment, n' is 5, m' is 1 and n'' is 3. In yet another embodiment, n' is 5, m' is 2 and n'' is 3. In yet another embodiment, n' is 5, m' is 3 and n'' is 3.

在一特定實施例中，n'為1，m'為0且n''為4。在一特定實施例中，n'為1，m'為1且n''為4。在一特定實施例中，n'為1，m'為2且n''為4。在一特定實施例中，n'為1，m'為3且n''為4。In a specific embodiment, n' is 1, m' is 0 and n'' is 4. In a specific embodiment, n' is 1, m' is 1 and n'' is 4. In a specific embodiment, n' is 1, m' is 2 and n'' is 4. In a specific embodiment, n' is 1, m' is 3 and n'' is 4.

在另一實施例中，n'為2，m'為0且n''為4。在另一實施例中，n'為2，m'為1且n''為4。在另一實施例中，n'為2，m'為2且n''為4。在另一實施例中，n'為2，m'為3且n''為4。In another embodiment, n' is 2, m' is 0 and n'' is 4. In another embodiment, n' is 2, m' is 1 and n'' is 4. In another embodiment, n' is 2, m' is 2 and n'' is 4. In another embodiment, n' is 2, m' is 3 and n'' is 4.

在又一實施例中，n'為3，m'為0且n''為4。在又一實施例中，n'為3，m'為1且n''為4。在又一實施例中，n'為3，m'為2且n''為4。在又一實施例中，n'為3，m'為3且n''為4。In yet another embodiment, n' is 3, m' is 0 and n'' is 4. In yet another embodiment, n' is 3, m' is 1 and n'' is 4. In yet another embodiment, n' is 3, m' is 2 and n'' is 4. In yet another embodiment, n' is 3, m' is 3 and n'' is 4.

在又另一實施例中，n'為4，m'為0且n''為4。在又另一實施例中，n'為4，m'為1且n''為4。在又另一實施例中，n'為4，m'為2且n''為4。在又另一實施例中，n'為4，m'為3且n''為4。In yet another embodiment, n' is 4, m' is 0 and n'' is 4. In yet another embodiment, n' is 4, m' is 1 and n'' is 4. In yet another embodiment, n' is 4, m' is 2 and n'' is 4. In yet another embodiment, n' is 4, m' is 3 and n'' is 4.

在又另一實施例中，n'為5，m'為0且n''為4。在又另一實施例中，n'為5，m'為1且n''為4。在又另一實施例中，n'為5，m'為2且n''為4。在又另一實施例中，n'為5，m'為3且n''為4。In yet another embodiment, n' is 5, m' is 0 and n'' is 4. In yet another embodiment, n' is 5, m' is 1 and n'' is 4. In yet another embodiment, n' is 5, m' is 2 and n'' is 4. In yet another embodiment, n' is 5, m' is 3 and n'' is 4.

在一特定實施例中，n'為1，m'為0且n''為5。在一特定實施例中，n'為1，m'為1且n''為5。在一特定實施例中，n'為1，m'為2且n''為5。在一特定實施例中，n'為1，m'為3且n''為5。In a specific embodiment, n' is 1, m' is 0 and n'' is 5. In a specific embodiment, n' is 1, m' is 1 and n'' is 5. In a specific embodiment, n' is 1, m' is 2 and n'' is 5. In a specific embodiment, n' is 1, m' is 3 and n'' is 5.

在另一實施例中，n'為2，m'為0且n''為5。在另一實施例中，n'為2，m'為1且n''為5。在另一實施例中，n'為2，m'為2且n''為5。在另一實施例中，n'為2，m'為3且n''為5。In another embodiment, n' is 2, m' is 0 and n'' is 5. In another embodiment, n' is 2, m' is 1 and n'' is 5. In another embodiment, n' is 2, m' is 2 and n'' is 5. In another embodiment, n' is 2, m' is 3 and n'' is 5.

在又一實施例中，n'為3，m'為0且n''為5。在又一實施例中，n'為3，m'為1且n''為5。在又一實施例中，n'為3，m'為2且n''為5。在又一實施例中，n'為3，m'為3且n''為5。In yet another embodiment, n' is 3, m' is 0 and n'' is 5. In yet another embodiment, n' is 3, m' is 1 and n'' is 5. In yet another embodiment, n' is 3, m' is 2 and n'' is 5. In yet another embodiment, n' is 3, m' is 3 and n'' is 5.

在又另一實施例中，n'為4，m'為0且n''為5。在又另一實施例中，n'為4，m'為1且n''為5。在又另一實施例中，n'為4，m'為2且n''為5。在又另一實施例中，n'為4，m'為3且n''為5。In yet another embodiment, n' is 4, m' is 0 and n'' is 5. In yet another embodiment, n' is 4, m' is 1 and n'' is 5. In yet another embodiment, n' is 4, m' is 2 and n'' is 5. In yet another embodiment, n' is 4, m' is 3 and n'' is 5.

在又另一實施例中，n'為5，m'為0且n''為5。在又另一實施例中，n'為5，m'為1且n''為5。在又另一實施例中，n'為5，m'為2且n''為5。在又另一實施例中，n'為5，m'為3且n''為5。In yet another embodiment, n' is 5, m' is 0 and n'' is 5. In yet another embodiment, n' is 5, m' is 1 and n'' is 5. In yet another embodiment, n' is 5, m' is 2 and n'' is 5. In yet another embodiment, n' is 5, m' is 3 and n'' is 5.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(XII)，其中X為O(CH 2CH 2O) mCH 2CH 2，其中m係選自1至4，且其中X'為CH 2(CH 2) n ''，其中n''係選自0至10。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (XII), wherein X is O(CH 2 CH 2 O) m CH 2 CH 2 , wherein m is selected from 1 to 4, and wherein X' is CH 2 (CH 2 ) n '' , wherein n'' is selected from 0 to 10.

在一實施例中，m係選自1至3且n''係選自0至10。在一實施例中，m係選自1至3且n''係選自0至5。在一實施例中，m係選自1至2且n''係選自0至3。在一實施例中，m係選自1至2且n''係選自0至2。在一特定實施例中，m為1且n''為0。在另一實施例中，m為2且n''為0。在又一實施例中，m為3且n''為0。在又另一實施例中，m為4且n''為0。在一特定實施例中，m為1且n''為1。在另一實施例中，m為2且n''為1。在又一實施例中，m為3且n''為1。在又另一實施例中，m為4且n''為1。在一特定實施例中，m為1且n''為2。在另一實施例中，m為2且n''為2。在又一實施例中，m為3且n''為2。在又另一實施例中，m為4且n''為2。在一特定實施例中，m為1且n''為3。在另一實施例中，m為2且n''為3。在又一實施例中，m為3且n''為3。在又另一實施例中，m為4且n''為3。在一特定實施例中，m為1且n''為4。在另一實施例中，m為2且n''為4。在又一實施例中，m為3且n''為4。在又另一實施例中，m為4且n''為4。在一特定實施例中，m為1且n''為5。在另一實施例中，m為2且n''為5。在又一實施例中，m為3且n''為5。在又另一實施例中，m為4且n''為5。在一特定實施例中，m為1且n''為6。在另一實施例中，m為2且n''為6。在又一實施例中，m為3且n''為6。在又另一實施例中，m為4且n''為6。In one embodiment, m is selected from 1 to 3 and n'' is selected from 0 to 10. In one embodiment, m is selected from 1 to 3 and n'' is selected from 0 to 5. In one embodiment, m is selected from 1 to 2 and n'' is selected from 0 to 3. In one embodiment, m is selected from 1 to 2 and n'' is selected from 0 to 2. In a specific embodiment, m is 1 and n'' is 0. In another embodiment, m is 2 and n'' is 0. In yet another embodiment, m is 3 and n'' is 0. In yet another embodiment, m is 4 and n'' is 0. In a specific embodiment, m is 1 and n'' is 1. In another embodiment, m is 2 and n'' is 1. In yet another embodiment, m is 3 and n'' is 1. In yet another embodiment, m is 4 and n'' is 1. In a particular embodiment, m is 1 and n'' is 2. In another embodiment, m is 2 and n'' is 2. In yet another embodiment, m is 3 and n'' is 2. In yet another embodiment, m is 4 and n'' is 2. In a particular embodiment, m is 1 and n'' is 3. In another embodiment, m is 2 and n'' is 3. In yet another embodiment, m is 3 and n'' is 3. In yet another embodiment, m is 4 and n'' is 3. In a particular embodiment, m is 1 and n'' is 4. In another embodiment, m is 2 and n'' is 4. In yet another embodiment, m is 3 and n'' is 4. In yet another embodiment, m is 4 and n'' is 4. In a particular embodiment, m is 1 and n'' is 5. In another embodiment, m is 2 and n" is 5. In yet another embodiment, m is 3 and n" is 5. In yet another embodiment, m is 4 and n" is 5. In a particular embodiment, m is 1 and n" is 6. In another embodiment, m is 2 and n" is 6. In yet another embodiment, m is 3 and n" is 6. In yet another embodiment, m is 4 and n" is 6.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(XII)，其中X為O(CH 2CH 2O) mCH 2CH 2，其中m係選自1至4，且其中X'為CH 2O(CH 2) n ''CH 2，其中n''係選自0至10。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (XII), wherein X is O(CH 2 CH 2 O) m CH 2 CH 2 , wherein m is selected from 1 to 4, and wherein X' is CH 2 O(CH 2 ) n '' CH 2 , wherein n'' is selected from 0 to 10.

在一實施例中，m係選自1至3且n''係選自0至10。在一實施例中，m係選自1至3且n''係選自0至5。在一實施例中，m係選自1至2且n''係選自0至3。在一實施例中，m係選自1至2且n''係選自0至2。在一特定實施例中，m為1且n''為0。在另一實施例中，m為2且n''為0。在又一實施例中，m為3且n''為0。在又另一實施例中，m為4且n''為0。在一特定實施例中，m為1且n''為1。在另一實施例中，m為2且n''為1。在又一實施例中，m為3且n''為1。在又另一實施例中，m為4且n''為1。在一特定實施例中，m為1且n''為2。在另一實施例中，m為2且n''為2。在又一實施例中，m為3且n''為2。在又另一實施例中，m為4且n''為2。在一特定實施例中，m為1且n''為3。在另一實施例中，m為2且n''為3。在又一實施例中，m為3且n''為3。在又另一實施例中，m為4且n''為3。在一特定實施例中，m為1且n''為4。在另一實施例中，m為2且n''為4。在又一實施例中，m為3且n''為4。在又另一實施例中，m為4且n''為4。在一特定實施例中，m為1且n''為5。在另一實施例中，m為2且n''為5。在又一實施例中，m為3且n''為5。在又另一實施例中，m為4且n''為5。在一特定實施例中，m為1且n''為6。在另一實施例中，m為2且n''為6。在又一實施例中，m為3且n''為6。在又另一實施例中，m為4且n''為6。In one embodiment, m is selected from 1 to 3 and n'' is selected from 0 to 10. In one embodiment, m is selected from 1 to 3 and n'' is selected from 0 to 5. In one embodiment, m is selected from 1 to 2 and n'' is selected from 0 to 3. In one embodiment, m is selected from 1 to 2 and n'' is selected from 0 to 2. In a specific embodiment, m is 1 and n'' is 0. In another embodiment, m is 2 and n'' is 0. In yet another embodiment, m is 3 and n'' is 0. In yet another embodiment, m is 4 and n'' is 0. In a specific embodiment, m is 1 and n'' is 1. In another embodiment, m is 2 and n'' is 1. In yet another embodiment, m is 3 and n'' is 1. In yet another embodiment, m is 4 and n'' is 1. In a particular embodiment, m is 1 and n'' is 2. In another embodiment, m is 2 and n'' is 2. In yet another embodiment, m is 3 and n'' is 2. In yet another embodiment, m is 4 and n'' is 2. In a particular embodiment, m is 1 and n'' is 3. In another embodiment, m is 2 and n'' is 3. In yet another embodiment, m is 3 and n'' is 3. In yet another embodiment, m is 4 and n'' is 3. In a particular embodiment, m is 1 and n'' is 4. In another embodiment, m is 2 and n'' is 4. In yet another embodiment, m is 3 and n'' is 4. In yet another embodiment, m is 4 and n'' is 4. In a particular embodiment, m is 1 and n'' is 5. In another embodiment, m is 2 and n" is 5. In yet another embodiment, m is 3 and n" is 5. In yet another embodiment, m is 4 and n" is 5. In a particular embodiment, m is 1 and n" is 6. In another embodiment, m is 2 and n" is 6. In yet another embodiment, m is 3 and n" is 6. In yet another embodiment, m is 4 and n" is 6.

在一實施例中，本發明提供一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(XII)，其中X為O(CH 2CH 2O) mCH 2CH 2，其中m係選自1至4，且其中X'為CH 2O(CH 2CH 2O) m '(CH 2) n ''CH 2，其中n''係選自0至10且m'係選自0至4。 In one embodiment, the present invention provides a serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (XII), wherein X is O(CH 2 CH 2 O) m CH 2 CH 2 , wherein m is selected from 1 to 4, and wherein X' is CH 2 O(CH 2 CH 2 O) m ' (CH 2 ) n '' CH 2 , wherein n'' is selected from 0 to 10 and m' is selected from 0 to 4.

在一實施例中，m係選自1至3，m'係選自0至4且n''係選自0至10。在一實施例中，m係選自1至2，m'係選自0至4且n''係選自0至5。在一實施例中，m係選自1至2，m'係選自0至2且n''係選自0至3。在一實施例中，m係選自1至2，m'係選自0至2且n''係選自0至1。In one embodiment, m is selected from 1 to 3, m' is selected from 0 to 4 and n'' is selected from 0 to 10. In one embodiment, m is selected from 1 to 2, m' is selected from 0 to 4 and n'' is selected from 0 to 5. In one embodiment, m is selected from 1 to 2, m' is selected from 0 to 2 and n'' is selected from 0 to 3. In one embodiment, m is selected from 1 to 2, m' is selected from 0 to 2 and n'' is selected from 0 to 1.

在一特定實施例中，m為1，m'為0且n''為0。在另一實施例中，m為1，m'為1且n''為0。在另一實施例中，m為1，m'為2且n''為0。在另一實施例中，m為1，m'為3且n''為0。In a particular embodiment, m is 1, m' is 0 and n'' is 0. In another embodiment, m is 1, m' is 1 and n'' is 0. In another embodiment, m is 1, m' is 2 and n'' is 0. In another embodiment, m is 1, m' is 3 and n'' is 0.

在另一實施例中，m為2，m'為0且n''為0。在另一實施例中，m為2，m'為1且n''為0。在另一實施例中，m為2，m'為2且n''為0。在另一實施例中，m為2，m'為3且n''為0。In another embodiment, m is 2, m' is 0 and n'' is 0. In another embodiment, m is 2, m' is 1 and n'' is 0. In another embodiment, m is 2, m' is 2 and n'' is 0. In another embodiment, m is 2, m' is 3 and n'' is 0.

在又一實施例中，m為3，m'為0且n''為0。在又一實施例中，m為3，m'為1且n''為0。在又一實施例中，m為3，m'為2且n''為0。在又一實施例中，m為3，m'為3且n''為0。In yet another embodiment, m is 3, m' is 0 and n'' is 0. In yet another embodiment, m is 3, m' is 1 and n'' is 0. In yet another embodiment, m is 3, m' is 2 and n'' is 0. In yet another embodiment, m is 3, m' is 3 and n'' is 0.

在又另一實施例中，m為4，m'為0且n''為0。在又另一實施例中，m為4，m'為1且n''為0。在又另一實施例中，m為4，m'為2且n''為0。在又另一實施例中，m為4，m'為3且n''為0。In yet another embodiment, m is 4, m' is 0 and n" is 0. In yet another embodiment, m is 4, m' is 1 and n" is 0. In yet another embodiment, m is 4, m' is 2 and n" is 0. In yet another embodiment, m is 4, m' is 3 and n" is 0.

在一特定實施例中，m為1，m'為0且n''為1。在一特定實施例中，m為1，m'為1且n''為1。在一特定實施例中，m為1，m'為2且n''為1。在一特定實施例中，m為1，m'為3且n''為1。In a specific embodiment, m is 1, m' is 0 and n'' is 1. In a specific embodiment, m is 1, m' is 1 and n'' is 1. In a specific embodiment, m is 1, m' is 2 and n'' is 1. In a specific embodiment, m is 1, m' is 3 and n'' is 1.

在另一實施例中，m為2，m'為0且n''為1。在另一實施例中，m為2，m'為1且n''為1。在另一實施例中，m為2，m'為2且n''為1。在另一實施例中，m為2，m'為3且n''為1。In another embodiment, m is 2, m' is 0 and n'' is 1. In another embodiment, m is 2, m' is 1 and n'' is 1. In another embodiment, m is 2, m' is 2 and n'' is 1. In another embodiment, m is 2, m' is 3 and n'' is 1.

在又一實施例中，m為3，m'為0且n''為1。在又一實施例中，m為3，m'為1且n''為1。在又一實施例中，m為3，m'為2且n''為1。在又一實施例中，m為3，m'為3且n''為1。In yet another embodiment, m is 3, m' is 0 and n'' is 1. In yet another embodiment, m is 3, m' is 1 and n'' is 1. In yet another embodiment, m is 3, m' is 2 and n'' is 1. In yet another embodiment, m is 3, m' is 3 and n'' is 1.

在又另一實施例中，m為4，m'為0且n''為1。在又另一實施例中，m為4，m'為1且n''為1。在又另一實施例中，m為4，m'為2且n''為1。在又另一實施例中，m為4，m'為3且n''為1。In yet another embodiment, m is 4, m' is 0 and n'' is 1. In yet another embodiment, m is 4, m' is 1 and n'' is 1. In yet another embodiment, m is 4, m' is 2 and n'' is 1. In yet another embodiment, m is 4, m' is 3 and n'' is 1.

在一特定實施例中，m為1，m'為0且n''為2。在一特定實施例中，m為1，m'為1且n''為2。在一特定實施例中，m為1，m'為2且n''為2。在一特定實施例中，m為1，m'為3且n''為2。In a specific embodiment, m is 1, m' is 0 and n'' is 2. In a specific embodiment, m is 1, m' is 1 and n'' is 2. In a specific embodiment, m is 1, m' is 2 and n'' is 2. In a specific embodiment, m is 1, m' is 3 and n'' is 2.

在另一實施例中，m為2，m'為0且n''為2。在另一實施例中，m為2，m'為1且n''為2。在另一實施例中，m為2，m'為2且n''為2。在另一實施例中，m為2，m'為3且n''為2。In another embodiment, m is 2, m' is 0 and n'' is 2. In another embodiment, m is 2, m' is 1 and n'' is 2. In another embodiment, m is 2, m' is 2 and n'' is 2. In another embodiment, m is 2, m' is 3 and n'' is 2.

在又一實施例中，m為3，m'為0且n''為2。在又一實施例中，m為3，m'為1且n''為2。在又一實施例中，m為3，m'為2且n''為2。在又一實施例中，m為3，m'為3且n''為2。In yet another embodiment, m is 3, m' is 0 and n'' is 2. In yet another embodiment, m is 3, m' is 1 and n'' is 2. In yet another embodiment, m is 3, m' is 2 and n'' is 2. In yet another embodiment, m is 3, m' is 3 and n'' is 2.

在又另一實施例中，m為4，m'為0且n''為2。在又另一實施例中，m為4，m'為1且n''為2。在又另一實施例中，m為4，m'為2且n''為2。在又另一實施例中，m為4，m'為3且n''為2。In yet another embodiment, m is 4, m' is 0 and n'' is 2. In yet another embodiment, m is 4, m' is 1 and n'' is 2. In yet another embodiment, m is 4, m' is 2 and n'' is 2. In yet another embodiment, m is 4, m' is 3 and n'' is 2.

在一特定實施例中，m為1，m'為0且n''為3。在一特定實施例中，m為1，m'為1且n''為3。在一特定實施例中，m為1，m'為2且n''為3。在一特定實施例中，m為1，m'為3且n''為3。In a specific embodiment, m is 1, m' is 0 and n'' is 3. In a specific embodiment, m is 1, m' is 1 and n'' is 3. In a specific embodiment, m is 1, m' is 2 and n'' is 3. In a specific embodiment, m is 1, m' is 3 and n'' is 3.

在另一實施例中，m為2，m'為0且n''為3。在另一實施例中，m為2，m'為1且n''為3。在另一實施例中，m為2，m'為2且n''為3。在另一實施例中，m為2，m'為3且n''為3。In another embodiment, m is 2, m' is 0 and n'' is 3. In another embodiment, m is 2, m' is 1 and n'' is 3. In another embodiment, m is 2, m' is 2 and n'' is 3. In another embodiment, m is 2, m' is 3 and n'' is 3.

在又一實施例中，m為3，m'為0且n''為3。在又一實施例中，m為3，m'為1且n''為3。在又一實施例中，m為3，m'為2且n''為3。在又一實施例中，m為3，m'為3且n''為3。In yet another embodiment, m is 3, m' is 0 and n'' is 3. In yet another embodiment, m is 3, m' is 1 and n'' is 3. In yet another embodiment, m is 3, m' is 2 and n'' is 3. In yet another embodiment, m is 3, m' is 3 and n'' is 3.

在又另一實施例中，m為4，m'為0且n''為3。在又另一實施例中，m為4，m'為1且n''為3。在又另一實施例中，m為4，m'為2且n''為3。在又另一實施例中，m為4，m'為3且n''為3。In yet another embodiment, m is 4, m' is 0 and n'' is 3. In yet another embodiment, m is 4, m' is 1 and n'' is 3. In yet another embodiment, m is 4, m' is 2 and n'' is 3. In yet another embodiment, m is 4, m' is 3 and n'' is 3.

在一特定實施例中，m為1，m'為0且n''為4。在一特定實施例中，m為1，m'為1且n''為4。在一特定實施例中，m為1，m'為2且n''為4。在一特定實施例中，m為1，m'為3且n''為4。In a specific embodiment, m is 1, m' is 0 and n'' is 4. In a specific embodiment, m is 1, m' is 1 and n'' is 4. In a specific embodiment, m is 1, m' is 2 and n'' is 4. In a specific embodiment, m is 1, m' is 3 and n'' is 4.

在另一實施例中，m為2，m'為0且n''為4。在另一實施例中，m為2，m'為1且n''為4。在另一實施例中，m為2，m'為2且n''為4。在另一實施例中，m為2，m'為3且n''為4。In another embodiment, m is 2, m' is 0 and n'' is 4. In another embodiment, m is 2, m' is 1 and n'' is 4. In another embodiment, m is 2, m' is 2 and n'' is 4. In another embodiment, m is 2, m' is 3 and n'' is 4.

在又一實施例中，m為3，m'為0且n''為4。在又一實施例中，m為3，m'為1且n''為4。在又一實施例中，m為3，m'為2且n''為4。在又一實施例中，m為3，m'為3且n''為4。In yet another embodiment, m is 3, m' is 0 and n'' is 4. In yet another embodiment, m is 3, m' is 1 and n'' is 4. In yet another embodiment, m is 3, m' is 2 and n'' is 4. In yet another embodiment, m is 3, m' is 3 and n'' is 4.

在又另一實施例中，m為4，m'為0且n''為4。在又另一實施例中，m為4，m'為1且n''為4。在又另一實施例中，m為4，m'為2且n''為4。在又另一實施例中，m為4，m'為3且n''為4。In yet another embodiment, m is 4, m' is 0 and n'' is 4. In yet another embodiment, m is 4, m' is 1 and n'' is 4. In yet another embodiment, m is 4, m' is 2 and n'' is 4. In yet another embodiment, m is 4, m' is 3 and n'' is 4.

在一特定實施例中，m為1，m'為0且n''為5。在一特定實施例中，m為1，m'為1且n''為5。在一特定實施例中，m為1，m'為2且n''為5。在一特定實施例中，m為1，m'為3且n''為5。In a specific embodiment, m is 1, m' is 0 and n'' is 5. In a specific embodiment, m is 1, m' is 1 and n'' is 5. In a specific embodiment, m is 1, m' is 2 and n'' is 5. In a specific embodiment, m is 1, m' is 3 and n'' is 5.

在另一實施例中，m為2，m'為0且n''為5。在另一實施例中，m為2，m'為1且n''為5。在另一實施例中，m為2，m'為2且n''為5。在另一實施例中，m為2，m'為3且n''為5。In another embodiment, m is 2, m' is 0 and n'' is 5. In another embodiment, m is 2, m' is 1 and n'' is 5. In another embodiment, m is 2, m' is 2 and n'' is 5. In another embodiment, m is 2, m' is 3 and n'' is 5.

在又一實施例中，m為3，m'為0且n''為5。在又一實施例中，m為3，m'為1且n''為5。在又一實施例中，m為3，m'為2且n''為5。在又一實施例中，m為3，m'為3且n''為5。In yet another embodiment, m is 3, m' is 0 and n'' is 5. In yet another embodiment, m is 3, m' is 1 and n'' is 5. In yet another embodiment, m is 3, m' is 2 and n'' is 5. In yet another embodiment, m is 3, m' is 3 and n'' is 5.

在又另一實施例中，m為4，m'為0且n''為5。在又另一實施例中，m為4，m'為1且n''為5。在又另一實施例中，m為4，m'為2且n''為5。在又另一實施例中，m為4，m'為3且n''為5。In yet another embodiment, m is 4, m' is 0 and n'' is 5. In yet another embodiment, m is 4, m' is 1 and n'' is 5. In yet another embodiment, m is 4, m' is 2 and n'' is 5. In yet another embodiment, m is 4, m' is 3 and n'' is 5.

在本發明之一實施例中，使用本章節之替代點擊化學製備本發明之血清型38醣結合物。本發明亦係關於製備如上文所揭露之血清型38醣結合物的方法。In one embodiment of the present invention, the serotype 38 saccharide conjugate of the present invention is prepared using the substitution click chemistry of this section. The present invention also relates to a method for preparing the serotype 38 saccharide conjugate as disclosed above.

在一實施例中，點擊化學可包含三個步驟，(a)使經分離血清型38醣與碳酸衍生物及炔烴連接子在非質子性溶劑中反應以產生經活化之炔基醣(醣的活化)，(b)使載體蛋白與帶有N-羥基丁二醯亞胺(NHS)部分和疊氮基之試劑反應，其中NHS部分與胺基反應以形成醯胺鍵，從而獲得疊氮基官能化載體蛋白(載體蛋白的活化)，(c)藉由Cu + 1介導之疊氮化物-炔烴環加成反應使步驟(a)之經活化之炔基醣與步驟(b)之經活化的疊氮基-載體蛋白反應以形成醣結合物。 In one embodiment, the click chemistry may comprise three steps, (a) reacting the isolated serotype 38 sugar with a carbonate derivative and an alkyne linker in an aprotic solvent to produce an activated alkynyl sugar (activation of the sugar), (b) reacting a carrier protein with a reagent having an N-hydroxysuccinimide (NHS) moiety and an azido group, wherein the NHS moiety reacts with an amine group to form an amide bond, thereby obtaining an azido-functionalized carrier protein (activation of the carrier protein), (c) reacting the activated alkynyl sugar of step (a) with the activated azido-carrier protein of step (b) by a Cu + 1- mediated azido-alkyne cycloaddition reaction to form a carbohydrate conjugate.

在步驟(a)之後，醣被稱為經活化，且在本文中被稱作「經活化之醣」或「經活化之炔基醣」。After step (a), the carbohydrate is said to be activated, and is referred to herein as an "activated carbohydrate" or "activated alkynyl carbohydrate."

在步驟(b)之後，載體被稱為經活化，且被稱作「經活化之載體」或「經活化之疊氮基-載體」。After step (b), the support is said to be activated and is referred to as "activated support" or "activated azido-support".

如上文所提及，在活化(a)之前，可將血清型38醣之尺寸設定為目標分子量(MW)範圍。因此，在一實施例中，經分離之血清型38醣在用碳酸衍生物及炔烴連接子活化之前經尺寸設定。在一實施例中，經分離之血清型38醣經尺寸設定為上文所定義之目標分子量(MW)範圍中之任一者。在一實施例中，經分離之血清型38醣在用碳酸衍生物及炔烴連接子活化之前並未經尺寸設定。As mentioned above, prior to activation (a), the serotype 38 sugars may be sized to a target molecular weight (MW) range. Thus, in one embodiment, the separated serotype 38 sugars are sized prior to activation with a carbonate derivative and an alkyne linker. In one embodiment, the separated serotype 38 sugars are sized to any of the target molecular weight (MW) ranges defined above. In one embodiment, the separated serotype 38 sugars are not sized prior to activation with a carbonate derivative and an alkyne linker.

在一實施例中，該碳酸衍生物係選自由以下組成之群：1,1'-羰基二咪唑(CDI)、1,1'-羰基-二-(1,2,4-三唑) (CDT)、碳酸二丁二醯亞胺酯(DSC)及氯甲酸N-羥基丁二醯亞胺酯。In one embodiment, the carbonic acid derivative is selected from the group consisting of 1,1'-carbonyldiimidazole (CDI), 1,1'-carbonyl-di-(1,2,4-triazole) (CDT), disuccinimidyl carbonate (DSC) and N-hydroxysuccinimidyl chloroformate.

在一實施例中，該碳酸衍生物為1,1'-羰基二咪唑(CDI)。在另一實施例中，該碳酸衍生物為1,1'-羰基-二-(1,2,4-三唑) (CDT)。在另一實施例中，該碳酸衍生物為碳酸二丁二醯亞胺酯(DSC)。在又另一實施例中，該碳酸衍生物為氯甲酸N-羥基丁二醯亞胺酯。In one embodiment, the carbonic acid derivative is 1,1'-carbonyldiimidazole (CDI). In another embodiment, the carbonic acid derivative is 1,1'-carbonyl-di-(1,2,4-triazole) (CDT). In another embodiment, the carbonic acid derivative is disuccinimidyl carbonate (DSC). In yet another embodiment, the carbonic acid derivative is N-hydroxysuccinimidyl chloroformate.

在一實施例中，該碳酸衍生物為1,1'-羰基二咪唑(CDI)或1,1'-羰基-二-(1,2,4-三唑) (CDT)。在一實施例中，該碳酸衍生物為1,1'-羰基二咪唑(CDI)。在一實施例中，該碳酸衍生物為1,1'-羰基-二-(1,2,4-三唑) (CDT)。較佳地，該碳酸衍生物為碳酸二丁二醯亞胺酯(DSC)。In one embodiment, the carbonic acid derivative is 1,1'-carbonyldiimidazole (CDI) or 1,1'-carbonyl-di-(1,2,4-triazole) (CDT). In one embodiment, the carbonic acid derivative is 1,1'-carbonyldiimidazole (CDI). In one embodiment, the carbonic acid derivative is 1,1'-carbonyl-di-(1,2,4-triazole) (CDT). Preferably, the carbonic acid derivative is disuccinimidyl carbonate (DSC).

在一實施例中，該炔烴連接子為式(XIV)之化合物， 其中X係選自由CH 2、CH 2(CH 2) n、(CH 2CH 2O) mCH 2CH 2、NHCO(CH 2) n、NHCO(CH 2CH 2O) mCH 2CH 2、OCH 2(CH 2) n及O(CH 2CH 2O) mCH 2CH 2組成之群；其中n係選自1至10且m係選自1至4。 In one embodiment, the alkyne linker is a compound of formula (XIV), wherein X is selected from the group consisting of CH2 , CH2 ( CH2 ) n , ( CH2CH2O ) mCH2CH2 , NHCO( CH2 ) n , NHCO ( CH2CH2O ) mCH2CH2 , OCH2 ( CH2 ) n and O( CH2CH2O ) mCH2CH2 ; wherein n is selected from 1-10 and m is selected from 1-4.

在一實施例中，該炔烴連接子為式(XIV)之化合物，其中X為CH 2。 In one embodiment, the alkyne linker is a compound of formula (XIV), wherein X is CH 2 .

在一實施例中，該炔烴連接子為式(XIV)之化合物，其中X為CH 2(CH 2) n，且n係選自1至10。在一實施例中，n係選自1至5。在一實施例中，n係選自1至4。在一實施例中，n係選自1至3。在一實施例中，n係選自1至2。在一特定實施例中，n為1。在另一實施例中，n為2。在又一實施例中，n為3。在又另一實施例中，n為4。在又另一實施例中，n為5。在又另一實施例中，n為6。在又另一實施例中，n為7。在又另一實施例中，n為8。在又另一實施例中，n為9。在又另一實施例中，n為10。 In one embodiment, the alkyne linker is a compound of formula (XIV), wherein X is CH2 ( CH2 ) n , and n is selected from 1 to 10. In one embodiment, n is selected from 1 to 5. In one embodiment, n is selected from 1 to 4. In one embodiment, n is selected from 1 to 3. In one embodiment, n is selected from 1 to 2. In a particular embodiment, n is 1. In another embodiment, n is 2. In another embodiment, n is 3. In yet another embodiment, n is 4. In yet another embodiment, n is 5. In yet another embodiment, n is 6. In yet another embodiment, n is 7. In yet another embodiment, n is 8. In yet another embodiment, n is 9. In yet another embodiment, n is 10.

在一實施例中，該炔烴連接子為式(XIV)之化合物，其中X為(CH 2CH 2O) mCH 2CH 2，其中m係選自1至4。在一實施例中，m係選自1至3。在一實施例中，m係選自1至2。在一特定實施例中，m為1。在另一實施例中，m為2。在又一實施例中，m為3。在又另一實施例中，m為4。 In one embodiment, the alkyne linker is a compound of formula (XIV), wherein X is ( CH2CH2O ) mCH2CH2 , wherein m is selected from 1 to 4. In one embodiment, m is selected from 1 to 3. In one embodiment, m is selected from 1 to 2. In a specific embodiment, m is 1. In another embodiment, m is 2. In yet another embodiment, m is 3. In yet another embodiment, m is 4.

在一實施例中，該炔烴連接子為式(XIV)之化合物，其中X為NHCO(CH 2) n，且n係選自1至10。在一實施例中，n係選自1至5。在一實施例中，n係選自1至4。在一實施例中，n係選自1至3。在一實施例中，n係選自1至2。在一特定實施例中，n為1。在另一實施例中，n為2。在又一實施例中，n為3。在又另一實施例中，n為4。在又另一實施例中，n為5。在又另一實施例中，n為6。在又另一實施例中，n為7。在又另一實施例中，n為8。在又另一實施例中，n為9。在又另一實施例中，n為10。 In one embodiment, the alkyne linker is a compound of formula (XIV), wherein X is NHCO(CH 2 ) n , and n is selected from 1 to 10. In one embodiment, n is selected from 1 to 5. In one embodiment, n is selected from 1 to 4. In one embodiment, n is selected from 1 to 3. In one embodiment, n is selected from 1 to 2. In a particular embodiment, n is 1. In another embodiment, n is 2. In another embodiment, n is 3. In yet another embodiment, n is 4. In yet another embodiment, n is 5. In yet another embodiment, n is 6. In yet another embodiment, n is 7. In yet another embodiment, n is 8. In yet another embodiment, n is 9. In yet another embodiment, n is 10.

在一實施例中，該炔烴連接子為式(XIV)之化合物，其中X為NHCO(CH 2CH 2O) mCH 2CH 2，其中m係選自1至4。在一實施例中，m係選自1至3。在一實施例中，m係選自1至2。在一特定實施例中，m為1。在另一實施例中，m為2。在又一實施例中，m為3。在又另一實施例中，m為4。 In one embodiment, the alkyne linker is a compound of formula (XIV), wherein X is NHCO(CH 2 CH 2 O) m CH 2 CH 2 , wherein m is selected from 1 to 4. In one embodiment, m is selected from 1 to 3. In one embodiment, m is selected from 1 to 2. In a specific embodiment, m is 1. In another embodiment, m is 2. In yet another embodiment, m is 3. In yet another embodiment, m is 4.

在一實施例中，該炔烴連接子為式(XIV)之化合物，其中X為OCH 2(CH 2) n，且n係選自1至10。在一實施例中，n係選自1至5。在一實施例中，n係選自1至4。在一實施例中，n係選自1至3。在一實施例中，n係選自1至2。在一特定實施例中，n為1。在另一實施例中，n為2。在又一實施例中，n為3。在又另一實施例中，n為4。在又另一實施例中，n為5。在又另一實施例中，n為6。在又另一實施例中，n為7。在又另一實施例中，n為8。在又另一實施例中，n為9。在又另一實施例中，n為10。 In one embodiment, the alkyne linker is a compound of formula (XIV), wherein X is OCH2 ( CH2 ) n , and n is selected from 1 to 10. In one embodiment, n is selected from 1 to 5. In one embodiment, n is selected from 1 to 4. In one embodiment, n is selected from 1 to 3. In one embodiment, n is selected from 1 to 2. In a particular embodiment, n is 1. In another embodiment, n is 2. In another embodiment, n is 3. In yet another embodiment, n is 4. In yet another embodiment, n is 5. In yet another embodiment, n is 6. In yet another embodiment, n is 7. In yet another embodiment, n is 8. In yet another embodiment, n is 9. In yet another embodiment, n is 10.

在一實施例中，該炔烴連接子為式(XIV)之化合物，其中X為O(CH 2CH 2O) mCH 2CH 2，其中m係選自1至4。在一實施例中，m係選自1至3。在一實施例中，m係選自1至2。在一特定實施例中，m為1。在另一實施例中，m為2。在又一實施例中，m為3。在又另一實施例中，m為4。 In one embodiment, the alkyne linker is a compound of formula (XIV), wherein X is O( CH2CH2O ) mCH2CH2 , wherein m is selected from 1 to 4. In one embodiment, m is selected from 1 to 3. In one embodiment, m is selected from 1 to 2. In a specific embodiment, m is 1. In another embodiment, m is 2. In yet another embodiment, m is 3. In yet another embodiment, m is 4.

在一實施例中，該炔烴連接子為式(XV)之化合物， In one embodiment, the alkyne linker is a compound of formula (XV),

因此，在一較佳實施例中，該炔烴連接子為炔丙胺。Therefore, in a preferred embodiment, the alkyne linker is propargylamine.

在一實施例中，該帶有N-羥基丁二醯亞胺(NHS)部分及疊氮基之試劑為式(XVI)之化合物， 其中X係選自由(CH 2) nCH 2C=O及(CH 2CH 2O) mCH 2CH 2=O組成之群，其中n係選自0至10且m係選自0至4。 In one embodiment, the reagent having an N-hydroxysuccinimide (NHS) moiety and an azido group is a compound of formula (XVI), wherein X is selected from the group consisting of (CH 2 ) n CH 2 C═O and (CH 2 CH 2 O) m CH 2 CH 2 ═O, wherein n is selected from 0-10 and m is selected from 0-4.

在一實施例中，該帶有N-羥基丁二醯亞胺(NHS)部分及疊氮基之試劑為式(XVI)之化合物，其中X為(CH 2) nCH 2C=O，其中n係選自0至10。在一實施例中，n係選自0至5。在一實施例中，n係選自0至4。在一實施例中，n係選自0至3。在一實施例中，n係選自0至2。在一特定實施例中，n為0。在一特定實施例中，n為1。在另一實施例中，n為2。在又一實施例中，n為3。在又另一實施例中，n為4。在又另一實施例中，n為5。在又另一實施例中，n為6。在又另一實施例中，n為7。在又另一實施例中，n為8。在又另一實施例中，n為9。在又另一實施例中，n為10。 In one embodiment, the reagent having an N-hydroxysuccinimide (NHS) moiety and an azido group is a compound of formula (XVI), wherein X is (CH 2 ) n CH 2 C═O, wherein n is selected from 0 to 10. In one embodiment, n is selected from 0 to 5. In one embodiment, n is selected from 0 to 4. In one embodiment, n is selected from 0 to 3. In one embodiment, n is selected from 0 to 2. In a specific embodiment, n is 0. In a specific embodiment, n is 1. In another embodiment, n is 2. In another embodiment, n is 3. In yet another embodiment, n is 4. In yet another embodiment, n is 5. In yet another embodiment, n is 6. In yet another embodiment, n is 7. In yet another embodiment, n is 8. In yet another embodiment, n is 9. In yet another embodiment, n is 10.

在一實施例中，該帶有N-羥基丁二醯亞胺(NHS)部分及疊氮基之試劑為式(XVI)之化合物，其中X為(CH 2CH 2O) mCH 2CH 2=O，其中m係選自0至4。在一實施例中，m係選自0至3。在一實施例中，m係選自0至2。在一實施例中，m係選自0至1。在一特定實施例中，m為0。在一特定實施例中，m為1。在另一實施例中，m為2。在又一實施例中，m為3。在又另一實施例中，m為4。 In one embodiment, the reagent having an NHS moiety and an azido group is a compound of formula (XVI), wherein X is (CH 2 CH 2 O) m CH 2 CH 2 ═O, wherein m is selected from 0 to 4. In one embodiment, m is selected from 0 to 3. In one embodiment, m is selected from 0 to 2. In one embodiment, m is selected from 0 to 1. In a specific embodiment, m is 0. In a specific embodiment, m is 1. In another embodiment, m is 2. In another embodiment, m is 3. In yet another embodiment, m is 4.

在一實施例中，該帶有N-羥基丁二醯亞胺(NHS)部分及疊氮基之試劑為式(XVII)之化合物： In one embodiment, the reagent having an N-hydroxysuccinimide (NHS) moiety and an azido group is a compound of formula (XVII):

因此，在一實施例中，該帶有N-羥基丁二醯亞胺(NHS)部分及疊氮基之試劑為2-疊氮基乙酸(2,5-二側氧基吡咯啶-1-基)酯。Therefore, in one embodiment, the reagent having an N-hydroxysuccinimide (NHS) moiety and an azido group is 2-azidoacetic acid (2,5-dioxopyrrolidin-1-yl) ester.

在一實施例中，步驟a)包含使醣與碳酸衍生物反應，隨後使經碳酸衍生物活化之醣與炔烴連接子在非質子性溶劑中反應以產生經活化之炔基醣。In one embodiment, step a) comprises reacting a sugar with a carbonic acid derivative, and then reacting the sugar activated by the carbonic acid derivative with an alkyne linker in an aprotic solvent to produce an activated alkynyl sugar.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量在0.01至10莫耳當量之間的量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the sugar with a carbonic acid derivative in an amount between 0.01 and 10 molar equivalents relative to the amount of sugar present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量在0.1至10莫耳當量之間的量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the sugar with a carbonic acid derivative in an amount between 0.1 and 10 molar equivalents relative to the amount of sugar present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量在0.5至5莫耳當量之間的量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the sugar with a carbonic acid derivative in an amount between 0.5 and 5 molar equivalents relative to the amount of sugar present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量在1至5莫耳當量之間的量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the sugar with a carbonic acid derivative in an amount between 1 and 5 molar equivalents relative to the amount of sugar present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量在2至5莫耳當量之間的量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the sugar with a carbonic acid derivative in an amount between 2 and 5 molar equivalents relative to the amount of sugar present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量在5至10莫耳當量之間的量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the sugar with a carbonic acid derivative in an amount between 5 and 10 molar equivalents relative to the amount of sugar present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量在0.1至5莫耳當量之間的量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the sugar with a carbonic acid derivative in an amount between 0.1 and 5 molar equivalents relative to the amount of sugar present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量在0.5至2莫耳當量之間的量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the sugar with a carbonic acid derivative in an amount between 0.5 and 2 molar equivalents relative to the amount of sugar present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量約0.01莫耳當量之量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the carbohydrate with a carbonic acid derivative in an amount of about 0.01 molar equivalent relative to the amount of carbohydrate present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量約0.1莫耳當量之量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the carbohydrate with a carbonic acid derivative in an amount of about 0.1 molar equivalent relative to the amount of carbohydrate present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量約0.2莫耳當量之量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the carbohydrate with a carbonic acid derivative in an amount of about 0.2 molar equivalents relative to the amount of carbohydrate present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量約0.5莫耳當量之量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the carbohydrate with a carbonic acid derivative in an amount of about 0.5 molar equivalent relative to the amount of carbohydrate present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量約1莫耳當量之量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the carbohydrate with a carbonic acid derivative in an amount of about 1 molar equivalent relative to the amount of carbohydrate present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量約2莫耳當量之量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the carbohydrate with a carbonic acid derivative in an amount of about 2 molar equivalents relative to the amount of carbohydrate present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量約5莫耳當量之量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the carbohydrate with a carbonic acid derivative in an amount of about 5 molar equivalents relative to the amount of carbohydrate present in the reaction mixture.

在一個實施例中，步驟a)包含使醣與相對於反應混合物中所存在之醣的量約10莫耳當量之量的碳酸衍生物反應。In one embodiment, step a) comprises reacting the carbohydrate with a carbonic acid derivative in an amount of about 10 molar equivalents relative to the amount of carbohydrate present in the reaction mixture.

在一實施例中，在步驟a)處，經分離醣與碳酸衍生物在非質子性溶劑中反應。In one embodiment, at step a), the separated sugar is reacted with a carbonic acid derivative in an aprotic solvent.

在一個實施例中，經分離醣與碳酸衍生物在包含二甲亞碸(DMSO)或二甲基甲醯胺(DMF)之溶液中反應。在一個實施例中，經分離醣與碳酸衍生物在包含二甲基甲醯胺(DMF)之溶液中反應。在一個實施例中，經分離醣與碳酸衍生物在包含二甲亞碸(DMSO)之溶液中反應。In one embodiment, the separated sugars and the carbonic acid derivatives are reacted in a solution comprising dimethyl sulfoxide (DMSO) or dimethyl formamide (DMF). In one embodiment, the separated sugars and the carbonic acid derivatives are reacted in a solution comprising dimethyl formamide (DMF). In one embodiment, the separated sugars and the carbonic acid derivatives are reacted in a solution comprising dimethyl sulfoxide (DMSO).

在一較佳實施例中，經分離醣與碳酸衍生物在包含二甲亞碸(DMSO)之溶液中反應。In a preferred embodiment, the separated sugar is reacted with a carbonic acid derivative in a solution containing dimethyl sulfoxide (DMSO).

在一個實施例中，經分離醣與碳酸衍生物在基本上由二甲亞碸(DMSO)或二甲基甲醯胺(DMF)組成之溶液中反應。在一個實施例中，經分離醣與碳酸衍生物在基本上由二甲基甲醯胺(DMF)組成之溶液中反應。在一個實施例中，經分離醣與碳酸衍生物在基本上由二甲亞碸(DMSO)組成之溶液中反應。In one embodiment, the separated sugars and the carbonic acid derivatives are reacted in a solution consisting essentially of dimethyl sulfoxide (DMSO) or dimethylformamide (DMF). In one embodiment, the separated sugars and the carbonic acid derivatives are reacted in a solution consisting essentially of dimethylformamide (DMF). In one embodiment, the separated sugars and the carbonic acid derivatives are reacted in a solution consisting essentially of dimethyl sulfoxide (DMSO).

在一實施例中，經分離醣與碳酸衍生物在基本上由二甲基乙醯胺組成之溶液中反應。在一實施例中，經分離醣與碳酸衍生物在基本上由N-甲基-2-吡咯啶酮組成之溶液中反應。在一實施例中，經分離醣與碳酸衍生物在基本上由六甲基磷醯胺(HMPA)組成之溶液中反應。In one embodiment, the separated sugars and the carbonic acid derivatives are reacted in a solution consisting essentially of dimethylacetamide. In one embodiment, the separated sugars and the carbonic acid derivatives are reacted in a solution consisting essentially of N-methyl-2-pyrrolidone. In one embodiment, the separated sugars and the carbonic acid derivatives are reacted in a solution consisting essentially of hexamethylphosphatamide (HMPA).

在一較佳實施例中，經分離醣與碳酸衍生物在基本上由二甲亞碸(DMSO)組成之溶液中反應。In a preferred embodiment, the separated sugar is reacted with the carbonic acid derivative in a solution consisting essentially of dimethyl sulfoxide (DMSO).

在一個實施例中，經分離醣與碳酸衍生物在二甲亞碸(DMSO)或二甲基甲醯胺(DMF)中反應。在一個實施例中，經分離醣與碳酸衍生物在二甲基甲醯胺(DMF)中反應。在一個實施例中，經分離醣與碳酸衍生物在二甲亞碸(DMSO)中反應。In one embodiment, the separated sugars are reacted with carbonic acid derivatives in dimethyl sulfoxide (DMSO) or dimethyl formamide (DMF). In one embodiment, the separated sugars are reacted with carbonic acid derivatives in dimethyl formamide (DMF). In one embodiment, the separated sugars are reacted with carbonic acid derivatives in dimethyl sulfoxide (DMSO).

在一實施例中，經分離醣與碳酸衍生物在二甲基乙醯胺中反應。在一實施例中，經分離醣與碳酸衍生物在N-甲基-2-吡咯啶酮中反應。在一實施例中，經分離醣與碳酸衍生物在六甲基磷醯胺(HMPA)中反應。In one embodiment, the separated sugars are reacted with carbonic acid derivatives in dimethylacetamide. In one embodiment, the separated sugars are reacted with carbonic acid derivatives in N-methyl-2-pyrrolidone. In one embodiment, the separated sugars are reacted with carbonic acid derivatives in hexamethylphosphatamide (HMPA).

在一較佳實施例中，經分離醣與CDI在二甲亞碸(DMSO)中反應。在一實施例中，經分離醣與CDI在無水DMSO中反應。In a preferred embodiment, the separated sugars are reacted with CDI in dimethyl sulfoxide (DMSO). In one embodiment, the separated sugars are reacted with CDI in anhydrous DMSO.

已出人意料地發現，使經分離醣與CDI在水分含量約0.1%至1% (v/v)之環境中反應能夠避免副反應。因此，在一個實施例中，經分離醣與CDI在包含0.1%至1% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含0.1%至0.8% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含0.1%至0.5% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含0.1%至0.4% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含0.1%至0.3% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含0.1%至0.2% (v/v)水之非質子性溶劑中反應。It has been unexpectedly discovered that reacting the separated sugars with CDI in an environment with a moisture content of about 0.1% to 1% (v/v) can avoid side reactions. Therefore, in one embodiment, the separated sugars and CDI are reacted in an aprotic solvent containing 0.1% to 1% (v/v) water. In one embodiment, the separated sugars and CDI are reacted in an aprotic solvent containing 0.1% to 0.8% (v/v) water. In one embodiment, the separated sugars and CDI are reacted in an aprotic solvent containing 0.1% to 0.5% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing 0.1% to 0.4% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing 0.1% to 0.3% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing 0.1% to 0.2% (v/v) water.

在一個實施例中，經分離醣與CDI在包含0.2%至1% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含0.2%至0.8% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含0.2%至0.5% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含0.2%至0.4% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含0.2%至0.3% (v/v)水之非質子性溶劑中反應。In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing 0.2% to 1% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing 0.2% to 0.8% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing 0.2% to 0.5% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing 0.2% to 0.4% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing 0.2% to 0.3% (v/v) water.

在一個實施例中，經分離醣與CDI在包含0.3%至0.8% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含0.3%至0.5% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含0.3%至0.4% (v/v)水之非質子性溶劑中反應。In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing 0.3% to 0.8% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing 0.3% to 0.5% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing 0.3% to 0.4% (v/v) water.

較佳地，在一個實施例中，經分離醣與CDI在包含0.1%至0.5% (v/v)水之非質子性溶劑中反應。Preferably, in one embodiment, the separated sugars are reacted with CDI in an aprotic solvent comprising 0.1% to 0.5% (v/v) water.

在一個實施例中，經分離醣與CDI在包含約0.1% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含約0.2% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含約0.3% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含約0.4% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含約0.5% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含約0.6% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含約0.7% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含約0.8% (v/v)水之非質子性溶劑中反應。在一個實施例中，經分離醣與CDI在包含約0.9% (v/v)水之非質子性溶劑中反應。In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing about 0.1% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing about 0.2% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing about 0.3% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing about 0.4% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing about 0.5% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing about 0.6% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing about 0.7% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing about 0.8% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in an aprotic solvent containing about 0.9% (v/v) water.

在一個實施例中，經分離醣與CDI在包含0.1%至1% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含0.1%至0.8% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含0.1%至0.5% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含0.1%至0.4% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含0.1%至0.3% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含0.1%至0.2% (v/v)水之DMSO中反應。In one embodiment, the separated sugars are reacted with CDI in DMSO containing 0.1% to 1% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in DMSO containing 0.1% to 0.8% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in DMSO containing 0.1% to 0.5% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in DMSO containing 0.1% to 0.4% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in DMSO containing 0.1% to 0.3% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in DMSO containing 0.1% to 0.2% (v/v) water.

在一個實施例中，經分離醣與CDI在包含0.2%至1% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含0.2%至0.8% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含0.2%至0.5% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含0.2%至0.4% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含0.2%至0.3% (v/v)水之DMSO中反應。In one embodiment, the separated sugars are reacted with CDI in DMSO containing 0.2% to 1% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in DMSO containing 0.2% to 0.8% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in DMSO containing 0.2% to 0.5% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in DMSO containing 0.2% to 0.4% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in DMSO containing 0.2% to 0.3% (v/v) water.

在一個實施例中，經分離醣與CDI在包含0.3%至0.8% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含0.3%至0.5% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含0.3%至0.4% (v/v)水之DMSO中反應。In one embodiment, the separated sugars are reacted with CDI in DMSO containing 0.3% to 0.8% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in DMSO containing 0.3% to 0.5% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in DMSO containing 0.3% to 0.4% (v/v) water.

較佳地，在一個實施例中，經分離醣與CDI在包含0.1%至0.5% (v/v)水之DMSO中反應。Preferably, in one embodiment, the separated sugars are reacted with CDI in DMSO containing 0.1% to 0.5% (v/v) water.

在一個實施例中，經分離醣與CDI在包含約0.1% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含約0.2% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含約0.3% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含約0.4% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含約0.5% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含約0.6% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含約0.7% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含約0.8% (v/v)水之DMSO中反應。在一個實施例中，經分離醣與CDI在包含約0.9% (v/v)水之DMSO中反應。In one embodiment, the separated sugars and CDI are reacted in DMSO containing about 0.1% (v/v) water. In one embodiment, the separated sugars and CDI are reacted in DMSO containing about 0.2% (v/v) water. In one embodiment, the separated sugars and CDI are reacted in DMSO containing about 0.3% (v/v) water. In one embodiment, the separated sugars and CDI are reacted in DMSO containing about 0.4% (v/v) water. In one embodiment, the separated sugars and CDI are reacted in DMSO containing about 0.5% (v/v) water. In one embodiment, the separated sugars and CDI are reacted in DMSO containing about 0.6% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in DMSO containing about 0.7% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in DMSO containing about 0.8% (v/v) water. In one embodiment, the separated sugars are reacted with CDI in DMSO containing about 0.9% (v/v) water.

在一個實施例中，在添加炔烴連接子之前藉由添加水來淬滅游離碳酸衍生物。水可使游離CDI不活化。In one embodiment, the free carbonic acid derivative is quenched by adding water before adding the alkyne linker. Water can inactivate the free CDI.

因此，在一實施例中，在碳酸衍生物活化之後添加水。在一實施例中，添加水以使混合物中之總水含量達到約1%至約10% (v/v)之間。在一實施例中，添加水以使混合物中之總水含量達到約1%至約5% (v/v)之間。在一實施例中，添加水以使混合物中之總水含量達到約1% (v/v)。在一實施例中，添加水以使混合物中之總水含量達到約2% (v/v)。在一實施例中，添加水以使混合物中之總水含量達到約5% (v/v)。Therefore, in one embodiment, water is added after the carbonic acid derivative is activated. In one embodiment, water is added to bring the total water content in the mixture to between about 1% and about 10% (v/v). In one embodiment, water is added to bring the total water content in the mixture to between about 1% and about 5% (v/v). In one embodiment, water is added to bring the total water content in the mixture to about 1% (v/v). In one embodiment, water is added to bring the total water content in the mixture to about 2% (v/v). In one embodiment, water is added to bring the total water content in the mixture to about 5% (v/v).

一旦醣已與碳酸衍生物反應且在最終用水淬滅碳酸衍生物之後，經碳酸衍生物活化之醣便與炔烴連接子反應。Once the sugar has been reacted with the carbonate derivative and after the carbonate derivative is finally quenched with water, the carbonate-activated sugar is reacted with the alkyne linker.

在一個實施例中，步驟a)進一步包含使經碳酸衍生物活化之醣與相對於經活化醣之多醣重複單元的量(RU之莫耳當量)在0.01至10莫耳當量之間的量的炔烴連接子反應。In one embodiment, step a) further comprises reacting the carbonate-activated sugar with an alkyne linker in an amount between 0.01 and 10 molar equivalents relative to the amount of polysaccharide repeating units (molar equivalents of RU) of the activated sugar.

在一個實施例中，步驟a)進一步包含使經碳酸衍生物活化之醣與相對於經活化醣之多醣重複單元的量在0.1至5莫耳當量之間的量的炔烴連接子反應。In one embodiment, step a) further comprises reacting the carbonate-activated sugar with an alkyne linker in an amount between 0.1 and 5 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一個實施例中，步驟a)進一步包含使經碳酸衍生物活化之醣與相對於經活化醣之多醣重複單元的量在0.1至5莫耳當量之間的量的炔烴連接子反應。In one embodiment, step a) further comprises reacting the carbonate-activated sugar with an alkyne linker in an amount between 0.1 and 5 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一個實施例中，步驟a)進一步包含使經碳酸衍生物活化之醣與相對於經活化醣之多醣重複單元的量在0.5至2莫耳當量之間的量的炔烴連接子反應。In one embodiment, step a) further comprises reacting the carbonate-activated sugar with an alkyne linker in an amount between 0.5 and 2 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一個實施例中，步驟a)進一步包含使經碳酸衍生物活化之醣與相對於經活化醣之多醣重複單元的量在1至5莫耳當量之間的量的炔烴連接子反應。In one embodiment, step a) further comprises reacting the carbonate-activated carbohydrate with an alkyne linker in an amount between 1 and 5 molar equivalents relative to the amount of polysaccharide repeating units of the activated carbohydrate.

在上述實施例中，該碳酸衍生物為CDI。在另一實施例中，該碳酸衍生物為CDT。在上述實施例中，該碳酸衍生物較佳為DSC。In the above embodiment, the carbonic acid derivative is CDI. In another embodiment, the carbonic acid derivative is CDT. In the above embodiment, the carbonic acid derivative is preferably DSC.

在一個實施例中，步驟a)之後經活化醣之活化程度在0.5至50%之間。炔基醣之活化程度定義為與炔烴連接子連接之重複單元的百分比。In one embodiment, the degree of activation of the activated sugar after step a) is between 0.5 and 50%. The degree of activation of the alkynyl sugar is defined as the percentage of repeat units attached to the alkyne linker.

在一個實施例中，步驟a)之後經活化醣之活化程度在1至30%之間。在另一實施例中，步驟a)之後經活化醣之活化程度在2至25%之間。在另一實施例中，步驟a)之後經活化醣之活化程度在3至20%之間。In one embodiment, the degree of activation of the activated sugar after step a) is between 1 and 30%. In another embodiment, the degree of activation of the activated sugar after step a) is between 2 and 25%. In another embodiment, the degree of activation of the activated sugar after step a) is between 3 and 20%.

在另一實施例中，步驟a)之後經活化醣之活化程度在3至15%之間。在另一實施例中，步驟a)之後經活化醣之活化程度在4至15%之間。在一實施例中，步驟a)之後經活化醣之活化程度在1至6%之間。In another embodiment, the degree of activation of the activated sugar after step a) is between 3 and 15%. In another embodiment, the degree of activation of the activated sugar after step a) is between 4 and 15%. In one embodiment, the degree of activation of the activated sugar after step a) is between 1 and 6%.

在一實施例中，步驟a)之後經活化醣之活化程度在3至6%之間。在一實施例中，步驟a)之後經活化醣之活化程度在10至15%之間。在一較佳實施例中，步驟a)之後經活化醣之活化程度在15至50%之間。In one embodiment, the degree of activation of the activated sugar after step a) is between 3 and 6%. In one embodiment, the degree of activation of the activated sugar after step a) is between 10 and 15%. In a preferred embodiment, the degree of activation of the activated sugar after step a) is between 15 and 50%.

在一實施例中，步驟a)之後經活化醣之活化程度為約1%。在一實施例中，步驟a)之後經活化醣之活化程度為約2%。在一實施例中，步驟a)之後經活化醣之活化程度為約3%。在一實施例中，步驟a)之後經活化醣之活化程度為約4%。在一實施例中，步驟a)之後經活化醣之活化程度為約5%。在一實施例中，步驟a)之後經活化醣之活化程度為約6%。在一實施例中，步驟a)之後經活化醣之活化程度為約7%。在一實施例中，步驟a)之後經活化醣之活化程度為約8%。在一實施例中，步驟a)之後經活化醣之活化程度為約9%。在一實施例中，步驟a)之後經活化醣之活化程度為約10%。在一實施例中，步驟a)之後經活化醣之活化程度為約11%。在一實施例中，步驟a)之後經活化醣之活化程度為約12%。在一實施例中，步驟a)之後經活化醣之活化程度為約13%。在一實施例中，步驟a)之後經活化醣之活化程度為約14%。在一實施例中，步驟a)之後經活化醣之活化程度為約15%。在一實施例中，步驟a)之後經活化醣之活化程度為約16%。在一實施例中，步驟a)之後經活化醣之活化程度為約17%。在一實施例中，步驟a)之後經活化醣之活化程度為約18%。在一實施例中，步驟a)之後經活化醣之活化程度為約19%。在一實施例中，步驟a)之後經活化醣之活化程度為約20%。In one embodiment, the degree of activation of the activated sugar after step a) is about 1%. In one embodiment, the degree of activation of the activated sugar after step a) is about 2%. In one embodiment, the degree of activation of the activated sugar after step a) is about 3%. In one embodiment, the degree of activation of the activated sugar after step a) is about 4%. In one embodiment, the degree of activation of the activated sugar after step a) is about 5%. In one embodiment, the degree of activation of the activated sugar after step a) is about 6%. In one embodiment, the degree of activation of the activated sugar after step a) is about 7%. In one embodiment, the degree of activation of the activated sugar after step a) is about 8%. In one embodiment, the degree of activation of the activated sugar after step a) is about 9%. In one embodiment, the degree of activation of the activated sugar after step a) is about 10%. In one embodiment, the degree of activation of the activated sugar after step a) is about 11%. In one embodiment, the degree of activation of the activated sugar after step a) is about 12%. In one embodiment, the degree of activation of the activated sugar after step a) is about 13%. In one embodiment, the degree of activation of the activated sugar after step a) is about 14%. In one embodiment, the degree of activation of the activated sugar after step a) is about 15%. In one embodiment, the degree of activation of the activated sugar after step a) is about 16%. In one embodiment, the degree of activation of the activated sugar after step a) is about 17%. In one embodiment, the degree of activation of the activated sugar after step a) is about 18%. In one embodiment, the degree of activation of the activated sugar after step a) is about 19%. In one embodiment, the degree of activation of the activated sugar after step a) is about 20%.

在一個實施例中，步驟b)包含使載體蛋白與相對於載體蛋白上之離胺酸0.1至10莫耳當量之量的帶有N-羥基丁二醯亞胺(NHS)部分及疊氮基之試劑反應。In one embodiment, step b) comprises reacting the carrier protein with 0.1 to 10 molar equivalents of a reagent having an N-hydroxysuccinimide (NHS) moiety and an azido group relative to the lysine on the carrier protein.

在一個實施例中，步驟b)包含使載體蛋白與相對於載體蛋白上之離胺酸0.5至10莫耳當量之量的帶有N-羥基丁二醯亞胺(NHS)部分及疊氮基之試劑反應。In one embodiment, step b) comprises reacting the carrier protein with a reagent having an N-hydroxysuccinimide (NHS) moiety and an azido group in an amount of 0.5 to 10 molar equivalents relative to the lysine on the carrier protein.

在一個實施例中，步驟b)包含使載體蛋白與相對於載體蛋白上之離胺酸1至5莫耳當量之量的帶有N-羥基丁二醯亞胺(NHS)部分及疊氮基之試劑反應。In one embodiment, step b) comprises reacting the carrier protein with 1 to 5 molar equivalents of a reagent having an N-hydroxysuccinimide (NHS) moiety and an azido group relative to the lysine on the carrier protein.

在一個實施例中，步驟b)包含使載體蛋白與相對於載體蛋白上之離胺酸2至5莫耳當量之量的帶有N-羥基丁二醯亞胺(NHS)部分及疊氮基之試劑反應。In one embodiment, step b) comprises reacting the carrier protein with 2 to 5 molar equivalents of a reagent having an N-hydroxysuccinimide (NHS) moiety and an azido group relative to the lysine on the carrier protein.

在一個實施例中，步驟b)包含使載體蛋白與相對於載體蛋白上之離胺酸約10莫耳當量之量的帶有N-羥基丁二醯亞胺(NHS)部分及疊氮基之試劑反應。In one embodiment, step b) comprises reacting the carrier protein with a reagent having an N-hydroxysuccinimide (NHS) moiety and an azido group in an amount of about 10 molar equivalents relative to the lysine on the carrier protein.

在一個實施例中，步驟b)包含使載體蛋白與相對於載體蛋白上之離胺酸約7.5莫耳當量之量的帶有N-羥基丁二醯亞胺(NHS)部分及疊氮基之試劑反應。In one embodiment, step b) comprises reacting the carrier protein with a reagent having an N-hydroxysuccinimide (NHS) moiety and an azido group in an amount of about 7.5 molar equivalents relative to the lysine on the carrier protein.

在一個實施例中，步驟b)包含使載體蛋白與相對於載體蛋白上之離胺酸約5莫耳當量之量的帶有N-羥基丁二醯亞胺(NHS)部分及疊氮基之試劑反應。In one embodiment, step b) comprises reacting the carrier protein with a reagent having an N-hydroxysuccinimide (NHS) moiety and an azido group in an amount of about 5 molar equivalents relative to the lysine on the carrier protein.

在一個實施例中，步驟b)包含使載體蛋白與相對於載體蛋白上之離胺酸約2莫耳當量之量的帶有N-羥基丁二醯亞胺(NHS)部分及疊氮基之試劑反應。In one embodiment, step b) comprises reacting the carrier protein with a reagent having an N-hydroxysuccinimide (NHS) moiety and an azido group in an amount of about 2 molar equivalents relative to the lysine on the carrier protein.

在一個實施例中，步驟b)包含使載體蛋白與相對於載體蛋白上之離胺酸約1莫耳當量之量的帶有N-羥基丁二醯亞胺(NHS)部分及疊氮基之試劑反應。In one embodiment, step b) comprises reacting the carrier protein with a reagent having an N-hydroxysuccinimide (NHS) moiety and an azido group in an amount of about 1 molar equivalent relative to the lysine on the carrier protein.

在一個實施例中，步驟b)包含使載體蛋白與相對於載體蛋白上之離胺酸約0.5莫耳當量之量的帶有N-羥基丁二醯亞胺(NHS)部分及疊氮基之試劑反應。In one embodiment, step b) comprises reacting the carrier protein with a reagent having an N-hydroxysuccinimide (NHS) moiety and an azido group in an amount of about 0.5 molar equivalent relative to the lysine on the carrier protein.

在一個實施例中，步驟b)包含使載體蛋白與相對於載體蛋白上之離胺酸約0.1莫耳當量之量的帶有N-羥基丁二醯亞胺(NHS)部分及疊氮基之試劑反應。In one embodiment, step b) comprises reacting the carrier protein with a reagent having an N-hydroxysuccinimide (NHS) moiety and an azido group in an amount of about 0.1 molar equivalent relative to the lysine on the carrier protein.

在一個實施例中，步驟b)之後經活化載體之活化程度在1與50之間。經活化載體之活化程度定義為載體蛋白中變得與帶有N-羥基丁二醯亞胺(NHS)部分及疊氮基之試劑連接的離胺酸殘基之數目。In one embodiment, the degree of activation of the activated carrier after step b) is between 1 and 50. The degree of activation of the activated carrier is defined as the number of lysine residues in the carrier protein that become linked to the reagent with an NHS moiety and an azido group.

在一實施例中，載體蛋白為CRM 197，其含有39個離胺酸殘基。在該實施例中，步驟b)之後經活化載體之活化程度可在1至30之間。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度在5至20之間。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度在9至18之間。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度在8至11之間。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度在15至20之間。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約5。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約6。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約7。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約8。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約9。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約10。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約11。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約12。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約13。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約14。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約15。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約16。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約17。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約18。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約19。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約20。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約21。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約22。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約23。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約24。在另一實施例中，步驟b)之後經活化載體(CRM 197)之活化程度為約25。 In one embodiment, the carrier protein is CRM 197 , which contains 39 lysine residues. In this embodiment, the degree of activation of the activated carrier after step b) may be between 1 and 30. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is between 5 and 20. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is between 9 and 18. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is between 8 and 11. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is between 15 and 20. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 5. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 6. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 7. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 8. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 9. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 10. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 11. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 12. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 13. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 14. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 15. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 16. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 17. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 18. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 19. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 20. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 21. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 22. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 23. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 24. In another embodiment, the degree of activation of the activated carrier (CRM 197 ) after step b) is about 25.

在一實施例中，載體蛋白為SCP或其片段。在該實施例中，步驟b)之後經活化載體之活化程度可在1至50之間。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度在5至50之間。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度在7至45之間。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度在5至15之間。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度在20至30之間。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度在30至50之間。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度在30至40之間。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度在10至40之間。在一較佳實施例中，步驟b)之後經活化載體(SCP)之活化程度在15至25之間。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約5。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約7。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約10。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約13。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約15。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約20。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約26。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約30。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約35。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約37。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約40。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約45。在另一實施例中，步驟b)之後經活化載體(SCP)之活化程度為約50。In one embodiment, the carrier protein is SCP or a fragment thereof. In this embodiment, the degree of activation of the activated carrier after step b) may be between 1 and 50. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is between 5 and 50. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is between 7 and 45. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is between 5 and 15. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is between 20 and 30. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is between 30 and 50. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is between 30 and 40. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is between 10 and 40. In a preferred embodiment, the degree of activation of the activated carrier (SCP) after step b) is between 15 and 25. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 5. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 7. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 10. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 13. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 15. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 20. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 26. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 30. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 35. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 37. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 40. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 45. In another embodiment, the degree of activation of the activated carrier (SCP) after step b) is about 50.

在一實施例中，載體蛋白為TT或其片段。在該實施例中，步驟b)之後經活化載體之活化程度可在1至30之間。在另一實施例中，步驟b)之後經活化載體(TT)之活化程度在5至25之間。在另一實施例中，步驟b)之後經活化載體(TT)之活化程度在7至25之間。在另一實施例中，步驟b)之後經活化載體(TT)之活化程度在10至20之間。在另一實施例中，步驟b)之後經活化載體(TT)之活化程度為約5。在另一實施例中，步驟b)之後經活化載體(TT)之活化程度為約7。在另一實施例中，步驟b)之後經活化載體(TT)之活化程度為約10。在另一實施例中，步驟b)之後經活化載體(TT)之活化程度為約12。在另一實施例中，步驟b)之後經活化載體(TT)之活化程度為約15。在另一實施例中，步驟b)之後經活化載體(TT)之活化程度為約20。在另一實施例中，步驟b)之後經活化載體(TT)之活化程度為約25。在另一實施例中，步驟b)之後經活化載體(TT)之活化程度為約30。In one embodiment, the carrier protein is TT or a fragment thereof. In this embodiment, the degree of activation of the activated carrier after step b) may be between 1 and 30. In another embodiment, the degree of activation of the activated carrier (TT) after step b) is between 5 and 25. In another embodiment, the degree of activation of the activated carrier (TT) after step b) is between 7 and 25. In another embodiment, the degree of activation of the activated carrier (TT) after step b) is between 10 and 20. In another embodiment, the degree of activation of the activated carrier (TT) after step b) is about 5. In another embodiment, the degree of activation of the activated carrier (TT) after step b) is about 7. In another embodiment, the degree of activation of the activated carrier (TT) after step b) is about 10. In another embodiment, the degree of activation of the activated carrier (TT) after step b) is about 12. In another embodiment, the degree of activation of the activated carrier (TT) after step b) is about 15. In another embodiment, the degree of activation of the activated carrier (TT) after step b) is about 20. In another embodiment, the degree of activation of the activated carrier (TT) after step b) is about 25. In another embodiment, the degree of activation of the activated carrier (TT) after step b) is about 30.

在一實施例中，結合反應c)係在水性緩衝液中進行。在一實施例中，結合反應c)係在水性緩衝液中在作為催化劑之銅(I)存在下進行。在一實施例中，結合反應c)係在水性緩衝液中在氧化劑及作為催化劑之銅(I)存在下進行。在一較佳實施例中，結合反應c)係在水性緩衝液中在作為催化劑之銅(I)及作為氧化劑之抗壞血酸鹽存在下進行。在一實施例中，可進一步添加THPTA (參(3-羥丙基三唑基甲基)胺)及胺基胍以防止蛋白質發生副反應。因此，在一較佳實施例中，結合反應c)係在水性緩衝液中在作為催化劑之銅(I)及作為氧化劑之抗壞血酸鹽存在下進行，其中反應混合物進一步包含THPTA (參(3-羥丙基三唑基甲基)胺)及胺基胍。In one embodiment, the binding reaction c) is carried out in an aqueous buffer. In one embodiment, the binding reaction c) is carried out in an aqueous buffer in the presence of copper (I) as a catalyst. In one embodiment, the binding reaction c) is carried out in an aqueous buffer in the presence of an oxidant and copper (I) as a catalyst. In a preferred embodiment, the binding reaction c) is carried out in an aqueous buffer in the presence of copper (I) as a catalyst and ascorbate as an oxidant. In one embodiment, THPTA (tris(3-hydroxypropyltriazolylmethyl)amine) and aminoguanidine may be further added to prevent side reactions of the protein. Therefore, in a preferred embodiment, the combination reaction c) is carried out in an aqueous buffer in the presence of copper (I) as a catalyst and ascorbate as an oxidant, wherein the reaction mixture further comprises THPTA (tris(3-hydroxypropyltriazolylmethyl)amine) and aminoguanidine.

在一實施例中，在步驟c)處經活化炔基醣與經活化疊氮基-載體之初始輸入比率(重量/重量)在0.1與3之間。在一實施例中，在步驟c)處經活化炔基醣與經活化疊氮基-載體之初始輸入比率(重量/重量)在0.5與2之間。在一實施例中，在步驟c)處經活化炔基醣與經活化疊氮基-載體之初始輸入比率(重量/重量)在0.6與1.5之間。在一實施例中，在步驟c)處經活化炔基醣與經活化疊氮基-載體之初始輸入比率(重量/重量)在0.8與1之間。在一較佳實施例中，在步驟c)處經活化炔基醣與經活化疊氮基-載體之初始輸入比率(重量/重量)在0.8與1之間。在一較佳實施例中，在步驟c)處經活化炔基醣與經活化疊氮基-載體之初始輸入比率(重量/重量)在0.8與1.2之間。在一實施例中，在步驟c)處經活化炔基醣與經活化疊氮基-載體之初始輸入比率(重量/重量)為約0.5。在一實施例中，在步驟c)處經活化炔基醣與經活化疊氮基-載體之初始輸入比率(重量/重量)為約0.6。在一實施例中，在步驟c)處經活化炔基醣與經活化疊氮基-載體之初始輸入比率(重量/重量)為約0.7。在一實施例中，在步驟c)處經活化炔基醣與經活化疊氮基-載體之初始輸入比率(重量/重量)為約0.8。在一實施例中，在步驟c)處經活化炔基醣與經活化疊氮基-載體之初始輸入比率(重量/重量)為約0.9。在一實施例中，在步驟c)處經活化炔基醣與經活化疊氮基-載體之初始輸入比率(重量/重量)為約1。在一實施例中，在步驟c)處經活化炔基醣與經活化疊氮基-載體之初始輸入比率(重量/重量)為約1.1。在一實施例中，在步驟c)處經活化炔基醣與經活化疊氮基-載體之初始輸入比率(重量/重量)為約1.2。在一實施例中，在步驟c)處經活化炔基醣與經活化疊氮基-載體之初始輸入比率(重量/重量)為約1.3。在一實施例中，在步驟c)處經活化炔基醣與經活化疊氮基-載體之初始輸入比率(重量/重量)為約1.4。在一實施例中，在步驟c)處經活化炔基醣與經活化疊氮基-載體之初始輸入比率(重量/重量)為約1.5。在一實施例中，在步驟c)處經活化炔基醣與經活化疊氮基-載體之初始輸入比率(重量/重量)為約1.6。在一實施例中，在步驟c)處經活化炔基醣與經活化疊氮基-載體之初始輸入比率(重量/重量)為約1.7。在一實施例中，在步驟c)處經活化炔基醣與經活化疊氮基-載體之初始輸入比率(重量/重量)為約1.8。在一實施例中，在步驟c)處經活化炔基醣與經活化疊氮基-載體之初始輸入比率(重量/重量)為約1.9。在一實施例中，在步驟c)處經活化炔基醣與經活化疊氮基-載體之初始輸入比率(重量/重量)為約2。In one embodiment, the initial input ratio (weight/weight) of activated alkynyl sugar to activated azido-carrier at step c) is between 0.1 and 3. In one embodiment, the initial input ratio (weight/weight) of activated alkynyl sugar to activated azido-carrier at step c) is between 0.5 and 2. In one embodiment, the initial input ratio (weight/weight) of activated alkynyl sugar to activated azido-carrier at step c) is between 0.6 and 1.5. In one embodiment, the initial input ratio (weight/weight) of activated alkynyl sugar to activated azido-carrier at step c) is between 0.8 and 1. In a preferred embodiment, the initial input ratio (weight/weight) of activated alkynyl sugar to activated azido-carrier at step c) is between 0.8 and 1. In a preferred embodiment, the initial input ratio (weight/weight) of activated alkynyl sugar to activated azido-carrier at step c) is between 0.8 and 1.2. In one embodiment, the initial input ratio (weight/weight) of activated alkynyl sugar to activated azido-carrier at step c) is about 0.5. In one embodiment, the initial input ratio (weight/weight) of activated alkynyl sugar to activated azido-carrier at step c) is about 0.6. In one embodiment, the initial input ratio (weight/weight) of activated alkynyl sugar to activated azido-carrier at step c) is about 0.7. In one embodiment, the initial input ratio (weight/weight) of activated alkynyl sugar to activated azido-carrier at step c) is about 0.8. In one embodiment, the initial input ratio (weight/weight) of activated alkynyl sugar to activated azido-carrier at step c) is about 0.9. In one embodiment, the initial input ratio (weight/weight) of activated alkynyl sugar to activated azido-carrier at step c) is about 1. In one embodiment, the initial input ratio (weight/weight) of activated alkynyl sugar to activated azido-carrier at step c) is about 1.1. In one embodiment, the initial input ratio (weight/weight) of activated alkynyl sugar to activated azido-carrier at step c) is about 1.2. In one embodiment, the initial input ratio (weight/weight) of activated alkynyl sugar to activated azido-carrier at step c) is about 1.3. In one embodiment, the initial input ratio (weight/weight) of activated alkynyl sugar to activated azido-carrier at step c) is about 1.4. In one embodiment, the initial input ratio (weight/weight) of activated alkynyl sugar to activated azido-carrier at step c) is about 1.5. In one embodiment, the initial input ratio (weight/weight) of activated alkynyl sugar to activated azido-carrier at step c) is about 1.6. In one embodiment, the initial input ratio (weight/weight) of activated alkynyl sugar to activated azido-carrier at step c) is about 1.7. In one embodiment, the initial input ratio (weight/weight) of activated alkynyl sugar to activated azido-carrier at step c) is about 1.8. In one embodiment, the initial input ratio (weight/weight) of activated alkynyl sugar to activated azido-carrier at step c) is about 1.9. In one embodiment, the initial input ratio (weight/weight) of activated alkynyl sugar to activated azido-carrier at step c) is about 2.

在單擊結合反應之後，未反應之炔烴基可仍存在於結合物中，可使用適合的炔烴基封端劑對此等未反應之炔烴基進行封端。在一個實施例中，此炔烴基封端劑為帶有疊氮基之試劑。After the single-strand conjugation reaction, unreacted alkynyl groups may still exist in the conjugate, and these unreacted alkynyl groups can be capped using a suitable alkynyl capping agent. In one embodiment, the alkynyl capping agent is a reagent with an azido group.

在一實施例中，該炔烴基封端劑為式(XVIII)之化合物， 其中X為(CH 2) n，其中n係選自1至15。 In one embodiment, the alkynyl end-capping agent is a compound of formula (XVIII), wherein X is (CH 2 ) n , wherein n is selected from 1-15.

在一個實施例中，此炔烴基封端劑為3-疊氮基-1-丙醇。In one embodiment, the alkynyl end-capping agent is 3-azido-1-propanol.

因此，在一實施例中，在步驟c)之後，該方法進一步包含用炔烴基封端劑對結合物中保留之未反應炔烴基進行封端的步驟。Therefore, in one embodiment, after step c), the method further comprises the step of capping the unreacted alkynyl groups remaining in the conjugate with an alkynyl capping agent.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量在0.05至20莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed with an amount of the capping agent between 0.05 and 20 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量在0.1至15莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed with an amount of the capping agent between 0.1 and 15 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量在0.5至10莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed with an amount of the capping agent between 0.5 and 10 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量在0.5至5莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed with an amount of the capping agent between 0.5 and 5 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量在0.5至2莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed with an amount of the capping agent between 0.5 and 2 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量在0.5至1莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed with an amount of the capping agent between 0.5 and 1 molar equivalent relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量在1至2莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed with an amount of the capping agent between 1 and 2 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量在1.5至2.5莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed with an amount of the capping agent between 1.5 and 2.5 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量約0.5莫耳當量之量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed using a capping agent in an amount of about 0.5 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量約1莫耳當量之量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed using a capping agent in an amount of about 1 molar equivalent relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量約1.5莫耳當量之量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed using an amount of the capping agent in an amount of about 1.5 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量約2莫耳當量之量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed using a capping agent in an amount of about 2 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量約2.5莫耳當量之量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed using an amount of the capping agent in an amount of about 2.5 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應炔烴基之封端係用相對於經活化醣之多醣重複單元的量約5莫耳當量之量的封端劑進行。In one embodiment, the capping of unreacted alkynyl groups is performed using a capping agent in an amount of about 5 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在單擊結合反應之後，結合物中可保留有未反應之疊氮基，可使用適合的疊氮基封端劑對此等未反應之疊氮基進行封端。因此，在一實施例中，在步驟c)之後，使用適合的疊氮基封端劑對結合物中未反應之疊氮基進行封端。在一個實施例中，此疊氮基封端劑為帶有炔烴基之試劑。在一個實施例中，此疊氮基封端劑為帶有末端炔烴之試劑。在一個實施例中，此疊氮基封端劑為帶有環炔烴之試劑。After the single-stranded conjugation reaction, unreacted azido groups may remain in the conjugate, and suitable azido group capping agents may be used to cap such unreacted azido groups. Therefore, in one embodiment, after step c), suitable azido group capping agents are used to cap the unreacted azido groups in the conjugate. In one embodiment, the azido group capping agent is a reagent with an alkynyl group. In one embodiment, the azido group capping agent is a reagent with a terminal alkynyl group. In one embodiment, the azido group capping agent is a reagent with a cycloalkynyl group.

在一實施例中，此疊氮基封端劑為式(XIX)之化合物， 其中X為(CH 2) n，其中n係選自1至15。 In one embodiment, the azido-capping agent is a compound of formula (XIX), wherein X is (CH 2 ) n , wherein n is selected from 1-15.

在一個實施例中，此疊氮基封端劑為炔丙醇。In one embodiment, the azido-capping agent is propargyl alcohol.

因此，在一實施例中，在步驟c)之後，該方法進一步包含用疊氮基封端劑對結合物中保留之未反應疊氮基進行封端的步驟。Therefore, in one embodiment, after step c), the method further comprises the step of capping the unreacted azido groups remaining in the conjugate with an azido capping agent.

在一實施例中，未反應疊氮基之封端係用相對於經活化醣之多醣重複單元的量在0.05至20莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted azido groups is performed with a capping agent in an amount between 0.05 and 20 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應疊氮基之封端係用相對於經活化醣之多醣重複單元的量在0.5至10莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted azido groups is performed with a capping agent in an amount between 0.5 and 10 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應疊氮基之封端係用相對於經活化醣之多醣重複單元的量在0.5至5莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted azido groups is performed with an amount of the capping agent between 0.5 and 5 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應疊氮基之封端係用相對於經活化醣之多醣重複單元的量在0.5至2莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted azido groups is performed with a capping agent in an amount between 0.5 and 2 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應疊氮基之封端係用相對於經活化醣之多醣重複單元的量在0.5至1莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted azido groups is performed with a capping agent in an amount between 0.5 and 1 molar equivalent relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應疊氮基之封端係用相對於經活化醣之多醣重複單元的量在1至2莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted azido groups is performed with an amount of the capping agent between 1 and 2 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應疊氮基之封端係用相對於經活化醣之多醣重複單元的量在0.75至1.5莫耳當量之間的量的封端劑進行。In one embodiment, the capping of unreacted azido groups is performed with an amount of the capping agent between 0.75 and 1.5 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應疊氮基之封端係用相對於經活化醣之多醣重複單元的量約1莫耳當量之量的封端劑進行。In one embodiment, the capping of unreacted azido groups is performed using a capping agent in an amount of about 1 molar equivalent relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應疊氮基之封端係用相對於經活化醣之多醣重複單元的量約1.5莫耳當量之量的封端劑進行。In one embodiment, the capping of unreacted azido groups is performed using a capping agent in an amount of about 1.5 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應疊氮基之封端係用相對於經活化醣之多醣重複單元的量約0.5莫耳當量之量的封端劑進行。In one embodiment, the capping of unreacted azido groups is performed using a capping agent in an amount of about 0.5 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在一實施例中，未反應疊氮基之封端係用相對於經活化醣之多醣重複單元的量約2莫耳當量之量的封端劑進行。In one embodiment, the capping of unreacted azido groups is performed using a capping agent in an amount of about 2 molar equivalents relative to the amount of polysaccharide repeating units of the activated sugar.

在與載體蛋白結合之後，可藉由熟習此項技術者已知之多種技術純化血清型38醣結合物(相對於醣-蛋白質結合物之量富集)。此等技術包括透析、濃縮/透濾操作、切向流過濾沉澱/溶離、管柱層析(DEAE或疏水性相互作用層析)及深層過濾。因此，在一個實施例中，用於產生本發明之醣結合物的方法包含在產生醣結合物之後對其進行純化的步驟。After conjugation to the carrier protein, the serotype 38 glycoconjugate can be purified (enriched relative to the amount of glyco-protein conjugate) by a variety of techniques known to those skilled in the art. Such techniques include dialysis, concentration/filtration procedures, tangential flow filtration precipitation/elution, column chromatography (DEAE or hydrophobic interaction chromatography) and deep filtration. Therefore, in one embodiment, the method for producing the glycoconjugate of the present invention comprises a step of purifying the glycoconjugate after it is produced.

在一態樣中，本發明提供根據本文所揭露之方法中之任一者產生的血清型38醣結合物。In one aspect, the invention provides a serotype 38 saccharide conjugate produced according to any of the methods disclosed herein.

2.4 載體蛋白醣結合物之組分為與血清型38醣結合之載體蛋白。術語「蛋白載體」或「載體蛋白」或「載體」可在本文中互換使用。載體蛋白應適於標準結合程序。 2.4 Carrier Protein The components of the glycoconjugate are carrier proteins conjugated to serotype 38 glycoconjugates. The terms "protein carrier" or "carrier protein" or "carrier" are used interchangeably herein. The carrier protein should be suitable for standard conjugation procedures.

在一較佳實施例中，血清型38醣結合物之載體蛋白係在由以下組成之群中選擇：DT (白喉類毒素)、TT (破傷風類毒素)或TT之片段C、CRM 197(白喉毒素之無毒但抗原一致之變異體)、其他DT突變體(諸如CRM 176、CRM 228、CRM 45(Uchida等人(1973) J. Biol. Chem. 218:3838-3844)、CRM 9、CRM 102、CRM 103或CRM 107；及Nicholls及Youle在Genetically Engineered Toxins, 編: Frankel, Maecel Dekker Inc. (1992)中描述之其他突變；Glu-148缺失或突變為Asp、Gln或Ser，及/或Ala 158缺失或突變為Gly，及美國專利第4,709,017號及第4,950,740號中所揭露之其他突變；至少一或多個殘基Lys 516、Lys 526、Phe 530及/或Lys 534之突變，及美國專利第5,917,017號及第6,455,673號中所揭露之其他突變；或美國專利第5,843,711號中所揭露之片段，肺炎鏈球菌性肺炎鏈球菌溶血素(ply) (Kuo等人(1995) Infect lmmun 63:2706-2713)包括以一些型式解毒之ply，例如dPLY-GMBS (WO 2004/081515、WO 2006/032499)或dPLY-formol、PhtX (包括PhtA、PhtB、PhtD、PhtE (PhtA、PhtB、PhtD或PhtE之序列揭露於WO 00/37105及WO 00/39299中)及Pht蛋白之融合物，例如PhtDE融合物、PhtBE融合物、Pht A-E (WO 01/98334、WO 03/054007、WO 2009/000826)、OMPC (腦膜炎球菌性外膜蛋白) (其通常提取自腦膜炎奈瑟氏菌血清群B (EP0372501))、PorB (來自腦膜炎奈瑟氏菌)、PD (流感嗜血桿菌蛋白D；參見例如EP0594610 B)，或其免疫功能等效物、合成肽(EP0378881、EP0427347)、熱休克蛋白(WO 93/17712、WO 94/03208)、百日咳蛋白(WO 98/58668、EP0471177)、細胞介素、淋巴激素、生長因子或激素(WO 91/01146)、人工蛋白質，包含來自各種病原體衍生抗原之多種人類CD4+ T細胞抗原決定基(Falugi等人(2001) Eur J Immunol 31:3816-3824)，諸如N19蛋白(Baraldoi等人(2004) Infect lmmun 72:4884-4887)、肺炎鏈球菌性表面蛋白PspA (WO 02/091998)、鐵攝取蛋白(WO 01/72337)、難養芽孢梭菌之毒素A或B (WO 00/61761)、轉鐵蛋白結合蛋白、肺炎鏈球菌性黏附蛋白(PsaA)、重組銅綠假單胞菌(Pseudomonas aeruginosa)外毒素A (尤其其無毒突變體(諸如具有麩胺酸553處之取代的外毒素A (Douglas等人(1987) J. Bacteriol. 169 (11):4967-4971))。其他蛋白，諸如卵白蛋白、匙孔螺血氰蛋白(keyhole limpet hemocyanin；KLH)、牛血清白蛋白(BSA)或結核菌素之純化蛋白衍生物(PPD)亦可用作載體蛋白。其他適合的載體蛋白包括不活化之細菌毒素，諸如霍亂類毒素(例如依WO 2004/083251中所描述)；大腸桿菌LT；大腸桿菌ST；及來自銅綠假單胞菌之外毒素A。另一適合的載體蛋白為來自鏈球菌之C5a肽酶(SCP)。另一適合載體蛋白質為根瘤菌抗生物素蛋白[aa 45-179J-GGGGSSS-SP1500- AAA-SP0785] (CP1) (WO2020056202)。另一適合載體蛋白為Rhavi-連接子-PdT(G294P)-連接子-SP0435 [aa 62-185]融合蛋白(SPP2)，參見WO2023039223。WO2020/056202及WO2023/039223以引用的方式併入。SPP2尤其描述於WO2023/039223之部分[0245]至[250]處。 In a preferred embodiment, the carrier protein of the serotype 38 glycoconjugate is selected from the group consisting of DT (diphtheria toxoid), TT (tetanus toxoid) or fragment C of TT, CRM 197 (a non-toxic but antigenically identical variant of diphtheria toxin), other DT mutants (such as CRM 176 , CRM 228 , CRM 45 (Uchida et al. (1973) J. Biol. Chem. 218:3838-3844), CRM 9 , CRM 102 , CRM 103 or CRM 107 ; and other mutations described by Nicholls and Youle in Genetically Engineered Toxins, ed.: Frankel, Maecel Dekker Inc. (1992); Glu-148 is deleted or mutated to Asp, Gln or Ser, and/or Ala 158 deletion or mutation to Gly, and other mutations disclosed in U.S. Patent Nos. 4,709,017 and 4,950,740; mutation of at least one or more residues Lys 516, Lys 526, Phe 530 and/or Lys 534, and other mutations disclosed in U.S. Patent Nos. 5,917,017 and 6,455,673; or fragments disclosed in U.S. Patent No. 5,843,711, pneumococcal pneumolysin (ply) (Kuo et al. (1995) Infect lmmun 63:2706-2713) includes some forms of detoxified ply, such as dPLY-GMBS (WO 2004/081515, WO 2006/032499) or dPLY-formol, PhtX (including PhtA, PhtB, PhtD, PhtE (the sequence of PhtA, PhtB, PhtD or PhtE is disclosed in WO 00/37105 and WO 00/39299) and a fusion of a Pht protein, such as PhtDE fusion, PhtBE fusion, Pht AE (WO 01/98334, WO 03/054007, WO 2009/000826), OMPC (meningococcal outer membrane protein) (which is usually extracted from Neisseria meningitidis serogroup B (EP0372501)), PorB (from Neisseria meningitidis), PD (Haemophilus influenzae protein D; see, for example, EP0594610 B), or immunologically functional equivalents thereof, synthetic peptides (EP0378881, EP0427347), heat shock proteins (WO 93/17712, WO 94/03208), pertussis proteins (WO 98/58668, EP0471177), interleukins, lymphokines, growth factors or hormones (WO 91/01146), artificial proteins, including a variety of human CD4+ T cell antigenic determinants from various pathogen-derived antigens (Falugi et al. (2001) Eur J Immunol 31:3816-3824), such as N19 protein (Baraldoi et al. (2004) Infect lmmun 72:4884-4887), pneumococcal surface protein PspA (WO 02/091998), iron uptake protein (WO 01/72337), toxin A or B of Clostridium fastidiosa (WO 00/61761), transferrin binding protein, pneumococcal adhesion protein (PsaA), recombinant Pseudomonas aeruginosa exotoxin A (especially its avirulent mutants (such as exotoxin A with a substitution at glutamine 553 (Douglas et al. (1987) J. Bacteriol. 169 (11): 4967-4971)). Other proteins, such as ovalbumin, keyhole limpet hemocyanin (KLH), bovine serum albumin (BSA) or purified protein derivative of tuberculin (PPD) can also be used as carrier proteins. Other suitable carrier proteins include inactivated bacterial toxins such as cholera toxoid (e.g., as described in WO 2004/083251); E. coli LT; E. coli ST; and exotoxin A from Pseudomonas aeruginosa. Another suitable carrier protein is C5a peptidase (SCP) from Streptococcus. Another suitable carrier protein is Rhizobium avidin [aa 45-179J-GGGGSSS-SP1500- AAA-SP0785] (CP1) (WO2020056202). Another suitable carrier protein is Rhavi-linker-PdT (G294P)-linker-SP0435 [aa 62-185] fusion protein (SPP2), see WO2023039223. WO2020/056202 and WO2023/039223 are incorporated by reference. SPP2 is described in particular at sections [0245] to [250] of WO2023/039223.

在一較佳實施例中，本發明之血清型38醣結合物之載體蛋白係選自由以下組成之群：TT、DT、DT突變體(諸如CRM 197)及來自鏈球菌之C5a肽酶(SCP)。 In a preferred embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is selected from the group consisting of TT, DT, DT mutants (such as CRM 197 ) and C5a peptidase (SCP) from Streptococcus.

在一實施例中，血清型38醣結合物之載體蛋白為DT  (白喉類毒素)。在另一實施例中，血清型38醣結合物之載體蛋白質為TT (破傷風類毒素)。In one embodiment, the carrier protein of the serotype 38 glycoconjugate is DT (diphtheria toxoid). In another embodiment, the carrier protein of the serotype 38 glycoconjugate is TT (tetanus toxoid).

在另一實施例中，血清型38醣結合物之載體蛋白為PD (流感嗜血桿菌蛋白D；參見例如EP0594610 B)。In another embodiment, the carrier protein of the serotype 38 glycoconjugate is PD (Haemophilus influenzae protein D; see, e.g., EP0594610 B).

在一較佳實施例中，血清型38醣結合物之載體蛋白為CRM 197或來自鏈球菌之C5a肽酶(SCP)。 In a preferred embodiment, the carrier protein of the serotype 38 glycoconjugate is CRM 197 or C5a peptidase (SCP) from Streptococcus.

在另一實施例中，血清型38醣結合物之載體蛋白為根瘤菌抗生物素蛋白[aa 45-179J-GGGGSSS-SP1500- AAA-SP0785] (CP1)。In another embodiment, the carrier protein of the serotype 38 saccharide conjugate is rhizobium avidin [aa 45-179J-GGGGSSS-SP1500-AAA-SP0785] (CP1).

在另一實施例中，血清型38醣結合物之載體蛋白為Rhavi-連接子-PdT(G294P)-連接子-SP0435 [aa 62-185]融合蛋白(SPP2)。在一個實施例中，該SPP2具有如WO2023/039223之SEQ ID NO: 19中所闡述之胺基酸序列。In another embodiment, the carrier protein of the serotype 38 glycoconjugate is a Rhavi-linker-PdT (G294P)-linker-SP0435 [aa 62-185] fusion protein (SPP2). In one embodiment, the SPP2 has an amino acid sequence as described in SEQ ID NO: 19 of WO2023/039223.

在一較佳實施例中，血清型38醣與CRM 197蛋白結合。CRM 197蛋白為一種無毒形式之白喉毒素，但在免疫學上與白喉毒素無法區分。CRM 197係由受無毒噬菌體β197 tox -感染之白喉棒狀桿菌(Corynebacterium diphtheriae)產生，該無毒噬菌體由產毒棒狀桿菌噬菌體β之亞硝基胍突變誘發產生(Uchida等人(1971) Nature New Biology 233:8-11)。CRM 197蛋白具有與白喉毒素相同的分子量，但因結構基因中之單個鹼基變化(鳥嘌呤變為腺嘌呤)而與白喉毒素不同。此單個鹼基變化引起成熟蛋白中之胺基酸取代(麩胺酸取代甘胺酸)，且消除白喉毒素之毒性特性。CRM 197蛋白為醣之安全且有效的T細胞依賴性載體。關於CRM 197及其產生之其他細節可例如見於美國專利第5,614,382號中。 In a preferred embodiment, the serotype 38 carbohydrate is conjugated to the CRM 197 protein. The CRM 197 protein is a non-toxic form of diphtheria toxin, but is immunologically indistinguishable from diphtheria toxin. CRM 197 is produced by Corynebacterium diphtheriae infected with the avirulent phage β197 tox - induced by nitrosoguanidine mutation of toxigenic coryneform phage β (Uchida et al. (1971) Nature New Biology 233:8-11). The CRM 197 protein has the same molecular weight as diphtheria toxin, but differs from diphtheria toxin by a single base change in the structural gene (guanine to adenine). This single base change results in an amino acid substitution (glutamine for glycine) in the mature protein and eliminates the toxic properties of diphtheria toxin. The CRM 197 protein is a safe and effective T cell-dependent carrier of sugars. Further details about CRM 197 and its production can be found, for example, in U.S. Patent No. 5,614,382.

在一實施例中，血清型38醣與CRM 197蛋白結合。在一實施例中，血清型38醣與CRM 197蛋白或CRM 197之A鏈結合(參見CN103495161)。在一實施例中，血清型38醣經由藉由重組大腸桿菌表現獲得之CRM 197之A鏈結合(參見CN103495161)。 In one embodiment, the serotype 38 saccharide is bound to the CRM 197 protein. In one embodiment, the serotype 38 saccharide is bound to the CRM 197 protein or the A chain of CRM 197 (see CN103495161). In one embodiment, the serotype 38 saccharide is bound via the A chain of CRM 197 expressed by recombinant E. coli (see CN103495161).

在其他較佳實施例中，本發明之血清型38醣結合物之載體蛋白為SCP (鏈球菌C5a肽酶)。In other preferred embodiments, the carrier protein of the serotype 38 glycoconjugate of the present invention is SCP (Streptococcus C5a peptidase).

β溶血性鏈球菌之兩種重要菌種化膿性鏈球菌(A組鏈球菌，GAS)及無乳鏈球菌(B組鏈球菌，GBS)引起多種嚴重人類感染，自輕症咽炎及膿皰至嚴重侵襲性疾病(諸如壞死性筋膜炎(GAS)及新生兒敗血症(GBS))都有，已針對此兩種菌種研發出消除此免疫反應之方式。β溶血性鏈球菌(包括GAS及GBS)之所有人類分離株均產生特異性地使C5a不活化之高度保守的細胞壁蛋白SCP (鏈球菌C5a肽酶)。來自GAS及GBS之scp基因編碼含有1,134至1,181個胺基酸之多肽(Brown等人, PNAS, 2005, 第102卷, 第51期, 第18391-18396頁)。前31個殘基為輸出信號前序列且在穿過細胞質膜時被移除。隨後68個殘基充當後序列且必須經移除以產生活性SCP。隨後10個殘基可經移除而不損失蛋白酶活性。在另一端，以Lys-1034開始為四個連續的17個殘基模體，接著為細胞分選及細胞壁連接信號。此組合信號由含有LPTTND序列之20個殘基親水性序列、17個殘基疏水性序列及較短鹼性羧基端構成。Two important species of beta-hemolytic streptococci, Streptococcus purulentus (Group A Streptococcus, GAS) and Streptococcus agalactiae (Group B Streptococcus, GBS), cause a variety of serious human infections, ranging from mild pharyngitis and abscesses to severe invasive diseases such as necrotizing fasciitis (GAS) and neonatal sepsis (GBS), and methods have been developed for these two species to eliminate this immune response. All human isolates of beta-hemolytic streptococci (including GAS and GBS) produce a highly conserved cell wall protein SCP (Streptococcal C5a peptidase) that specifically inactivates C5a. The scp genes from GAS and GBS encode polypeptides containing 1,134 to 1,181 amino acids (Brown et al., PNAS, 2005, Vol. 102, No. 51, pp. 18391-18396). The first 31 residues are the pre-sequence of the export signal and are removed when crossing the cytoplasmic membrane. The next 68 residues serve as the post-sequence and must be removed to produce active SCP. The next 10 residues can be removed without loss of protease activity. At the other end, there are four consecutive 17 residue motifs starting with Lys-1034, followed by cell sorting and cell wall attachment signals. This combined signal consists of a 20-residue hydrophilic sequence containing the LPTTND sequence, a 17-residue hydrophobic sequence, and a short alkaline carboxyl terminus.

SCP可分成域(參見Brown等人, PNAS, 2005, 第102卷, 第51號。第18391-18396頁之圖1B)。此等域為前域/後域(其包含輸出訊號前序列(通常為前31個殘基)及後序列(通常為後68個殘基))、蛋白酶域(其分解為兩部分(蛋白酶部分1，通常為殘基89-333/334；及蛋白酶域部分2，且通常為殘基467/468-583/584)、蛋白酶相關域(PA域) (通常為殘基333/334-467/468)、三個纖維結合蛋白質III型(Fn)域(Fn1，通常為殘基583/584-712/713；Fn2，通常為殘基712/713-928/929/930；通常Fn3，殘基929/930-1029/1030/1031)及細胞壁錨定域(通常為殘基1029/1030/1031至C端)。SCP can be divided into domains (see Brown et al., PNAS, 2005, Vol. 102, No. 51. Figure 1B on pages 18391-18396). These domains are the pro/post domain (which includes the pre-sequence of the output signal (usually the first 31 residues) and the post sequence (usually the last 68 residues)), the protease domain (which is divided into two parts (protease part 1, usually residues 89-333/334; and protease domain part 2, and usually residues 467/468-583/584), the protease-associated domain (PA domain) (usually residues 333/334-467/468), three fiber binding protein type III (Fn) domains (Fn1, usually residues 583/584-712/713; Fn2, usually residues 712/713-928/929/930; usually Fn3, residues 929/930-1029/1030/1031) and a cell wall anchoring domain (usually residues 1029/1030/1031 to the C-terminus).

在一實施例中，本發明之血清型38醣結合物之載體蛋白為來自GBS之SCP (SCPB)。SCPB之實例提供於WO97/26008之SEQ. ID.NO: 3。亦參見WO00/34487之SEQ ID NO: 3。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is SCP from GBS (SCPB). An example of SCPB is provided in SEQ. ID. NO: 3 of WO97/26008. See also SEQ ID NO: 3 of WO00/34487.

在另一較佳實施例中，本發明之血清型38醣結合物之載體蛋白為來自GAS之SCP (SCPA)。In another preferred embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is SCP (SCPA) from GAS.

SCPA之實例可見於WO97/26008之SEQ.ID.No.1及SEQ.ID.No.2。亦參見WO00/34487之SEQ ID NO: 1、2及23。Examples of SCPA can be found in SEQ.ID.No.1 and SEQ.ID.No.2 of WO97/26008. See also SEQ ID NOs: 1, 2 and 23 of WO00/34487.

在一較佳實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化之SCP。In a preferred embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is enzymatically inactive SCP.

在其他較佳實施例中，本發明之血清型38醣結合物之載體蛋白為來自GBS之酶不活化SCP (SCPB)。In other preferred embodiments, the carrier protein of the serotype 38 glycoconjugate of the present invention is enzymatically inactive SCP (SCPB) from GBS.

在另一較佳實施例中，本發明之血清型38醣結合物之載體蛋白為來自GAS之酶不活化SCP (SCPA)。In another preferred embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is enzymatically inactive SCP (SCPA) from GAS.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為SCP之片段。在一實施例中，本發明之血清型38醣結合物之載體蛋白為SCPA之片段。較佳地，本發明之血清型38醣結合物之載體蛋白為SCPB之片段。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is a fragment of SCP. In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is a fragment of SCPA. Preferably, the carrier protein of the serotype 38 glycoconjugate of the present invention is a fragment of SCPB.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為SCP之片段，其包含蛋白酶域、蛋白酶相關域(PA域)及三個纖維結合蛋白III型(Fn)域，但不包含輸出信號前序列、後序列及細胞壁錨定域。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is a fragment of SCP, which comprises a protease domain, a protease-associated domain (PA domain) and three fiber binding protein type III (Fn) domains, but does not comprise an export signal pre-sequence, a post-sequence and a cell wall anchoring domain.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為SCP之片段，其包含蛋白酶域、蛋白酶相關域(PA域)及三個纖維結合蛋白III型(Fn)域，但不包含輸出信號前序列、後序列及細胞壁錨定域。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is a fragment of SCP, which comprises a protease domain, a protease-associated domain (PA domain) and three fiber binding protein type III (Fn) domains, but does not comprise an export signal pre-sequence, a post-sequence and a cell wall anchoring domain.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為SCP之酶不活化片段，其包含蛋白酶域、蛋白酶相關域(PA域)以及三個纖維結合蛋白III型(Fn)域中之兩者，但不包含輸出信號前序列、後序列及細胞壁錨定域。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzymatically inactive fragment of SCP, which comprises a protease domain, a protease-associated domain (PA domain) and two of the three fiber binding protein type III (Fn) domains, but does not comprise an export signal pre-sequence, a post-sequence and a cell wall anchoring domain.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為SCP之酶不活化片段。在一實施例中，該SCP之酶不活化片段包含蛋白酶域、蛋白酶相關域(PA域)及三個纖維結合蛋白III型(Fn)域，但不包含輸出信號前序列、後序列及細胞壁錨定域。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzymatically inactive fragment of SCP. In one embodiment, the enzymatically inactive fragment of SCP comprises a protease domain, a protease-associated domain (PA domain) and three fiber binding protein type III (Fn) domains, but does not comprise an export signal pre-sequence, a post-sequence and a cell wall anchoring domain.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為SCPA之酶不活化片段。在一實施例中，該酶不活化的SCPA之片段包含蛋白酶域、蛋白酶相關域(PA域)及三個纖維結合蛋白III型(Fn)域，但不包含輸出信號前序列、後序列及細胞壁錨定域。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzymatically inactive fragment of SCPA. In one embodiment, the enzymatically inactive fragment of SCPA comprises a protease domain, a protease-associated domain (PA domain) and three fiber binding protein type III (Fn) domains, but does not comprise an export signal pre-sequence, a post-sequence and a cell wall anchoring domain.

在一較佳實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCPB之片段。較佳地，該酶不活化的SCPB之片段包含蛋白酶域、蛋白酶相關域(PA域)及三個纖維結合蛋白III型(Fn)域，但不包含輸出信號前序列、後序列及細胞壁錨定域。In a preferred embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzymatically inactive fragment of SCPB. Preferably, the enzymatically inactive fragment of SCPB comprises a protease domain, a protease-associated domain (PA domain) and three fiber binding protein type III (Fn) domains, but does not comprise an export signal pre-sequence, a post-sequence and a cell wall anchoring domain.

在一實施例中，SCP之酶活性藉由置換野生型序列之至少一個胺基酸而不活化。在一實施例中，該置換係選自由D130A、H193A、N295A及S512A組成之群。數目指示根據WO00/34487之SEQ ID NO: 1之編號的肽酶中之胺基酸殘基位置。In one embodiment, the enzymatic activity of SCP is inactivated by replacing at least one amino acid of the wild-type sequence. In one embodiment, the replacement is selected from the group consisting of D130A, H193A, N295A and S512A. The numbers indicate the amino acid residue positions in the peptidase according to the numbering of SEQ ID NO: 1 of WO00/34487.

因此，在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCP，其中該不活化係藉由置換野生型序列之至少一個胺基酸來實現。較佳地，至少一個胺基酸之該置換係在蛋白酶域中。在一實施例中，至少一個胺基酸之該置換係在蛋白酶域之部分1中。在一實施例中，至少一個胺基酸之該置換係在蛋白酶域之部分2中。在一實施例中，該置換係選自由D130A、H193A、N295A及S512A組成之群。在一實施例中，該置換為D130A。在另一實施例中，該置換為H193A。在另一實施例中，該置換為N295A。在又一實施例中，該置換為S512A。Therefore, in one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzyme-inactive SCP, wherein the inactivation is achieved by replacing at least one amino acid of the wild-type sequence. Preferably, the replacement of at least one amino acid is in the protease domain. In one embodiment, the replacement of at least one amino acid is in part 1 of the protease domain. In one embodiment, the replacement of at least one amino acid is in part 2 of the protease domain. In one embodiment, the replacement is selected from the group consisting of D130A, H193A, N295A and S512A. In one embodiment, the replacement is D130A. In another embodiment, the replacement is H193A. In another embodiment, the replacement is N295A. In yet another embodiment, the replacement is S512A.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCPA，其中該不活化係藉由置換野生型序列之至少一個胺基酸來實現。較佳地，至少一個胺基酸之該置換係在蛋白酶域中。在一實施例中，至少一個胺基酸之該置換係在蛋白酶域之部分1中。在一實施例中，至少一個胺基酸之該置換係在蛋白酶域之部分2中。在一實施例中，該置換係選自由D130A、H193A、N295A及S512A組成之群。在一實施例中，該置換為D130A。在另一實施例中，該置換為H193A。在另一實施例中，該置換為N295A。在又一實施例中，該置換為S512A。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzyme-inactive SCPA, wherein the inactivation is achieved by replacing at least one amino acid of the wild-type sequence. Preferably, the replacement of at least one amino acid is in the protease domain. In one embodiment, the replacement of at least one amino acid is in part 1 of the protease domain. In one embodiment, the replacement of at least one amino acid is in part 2 of the protease domain. In one embodiment, the replacement is selected from the group consisting of D130A, H193A, N295A and S512A. In one embodiment, the replacement is D130A. In another embodiment, the replacement is H193A. In another embodiment, the replacement is N295A. In yet another embodiment, the replacement is S512A.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCPB，其中該不活化係藉由置換野生型序列之至少一個胺基酸來實現。較佳地，至少一個胺基酸之該置換係在蛋白酶域中。在一實施例中，至少一個胺基酸之該置換係在蛋白酶域之部分1中。在一實施例中，至少一個胺基酸之該置換係在蛋白酶域之部分2中。在一實施例中，該置換係選自由D130A、H193A、N295A及S512A組成之群。在一實施例中，該置換為D130A。在另一實施例中，該置換為H193A。在另一實施例中，該置換為N295A。在又一實施例中，該置換為S512A。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzyme-inactive SCPB, wherein the inactivation is achieved by replacing at least one amino acid of the wild-type sequence. Preferably, the replacement of at least one amino acid is in the protease domain. In one embodiment, the replacement of at least one amino acid is in part 1 of the protease domain. In one embodiment, the replacement of at least one amino acid is in part 2 of the protease domain. In one embodiment, the replacement is selected from the group consisting of D130A, H193A, N295A and S512A. In one embodiment, the replacement is D130A. In another embodiment, the replacement is H193A. In another embodiment, the replacement is N295A. In yet another embodiment, the replacement is S512A.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCP之片段，其中該不活化係藉由置換野生型序列之至少一個胺基酸來實現。較佳地，至少一個胺基酸之該置換係在蛋白酶域中。在一實施例中，至少一個胺基酸之該置換係在蛋白酶域之部分1中。在一實施例中，至少一個胺基酸之該置換係在蛋白酶域之部分2中。在一實施例中，該置換係選自由D130A、H193A、N295A及S512A組成之群。在一實施例中，該置換為D130A。在另一實施例中，該置換為H193A。在另一實施例中，該置換為N295A。在又一實施例中，該置換為S512A。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is a fragment of an enzymatically inactive SCP, wherein the inactivation is achieved by replacing at least one amino acid of the wild-type sequence. Preferably, the replacement of at least one amino acid is in the protease domain. In one embodiment, the replacement of at least one amino acid is in part 1 of the protease domain. In one embodiment, the replacement of at least one amino acid is in part 2 of the protease domain. In one embodiment, the replacement is selected from the group consisting of D130A, H193A, N295A and S512A. In one embodiment, the replacement is D130A. In another embodiment, the replacement is H193A. In another embodiment, the replacement is N295A. In yet another embodiment, the replacement is S512A.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCP之片段，其包含蛋白酶域、蛋白酶相關域(PA域)及三個纖維結合蛋白III型(Fn)域但不包含輸出信號前序列、後序列及細胞壁錨定域，其中該不活化係藉由置換野生型序列之至少一個胺基酸來實現。較佳地，至少一個胺基酸之該置換係在蛋白酶域中。在一實施例中，至少一個胺基酸之該置換係在蛋白酶域之部分1中。在一實施例中，至少一個胺基酸之該置換係在蛋白酶域之部分2中。在一實施例中，該置換係選自由D130A、H193A、N295A及S512A組成之群。在一實施例中，該置換為D130A。在另一實施例中，該置換為H193A。在另一實施例中，該置換為N295A。在又一實施例中，該置換為S512A。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is a fragment of an enzyme-inactive SCP, which comprises a protease domain, a protease-associated domain (PA domain) and three fiber binding protein type III (Fn) domains but does not comprise an export signal presequence, a postsequence and a cell wall anchoring domain, wherein the inactivation is achieved by replacing at least one amino acid of the wild-type sequence. Preferably, the replacement of at least one amino acid is in the protease domain. In one embodiment, the replacement of at least one amino acid is in part 1 of the protease domain. In one embodiment, the replacement of at least one amino acid is in part 2 of the protease domain. In one embodiment, the replacement is selected from the group consisting of D130A, H193A, N295A and S512A. In one embodiment, the replacement is D130A. In another embodiment, the substitution is H193A. In another embodiment, the substitution is N295A. In yet another embodiment, the substitution is S512A.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCPA之片段，其包含蛋白酶域、蛋白酶相關域(PA域)及三個纖維結合蛋白III型(Fn)域但不包含輸出信號前序列、後序列及細胞壁錨定域，其中該不活化係藉由置換野生型序列之至少一個胺基酸來實現。較佳地，至少一個胺基酸之該置換係在蛋白酶域中。在一實施例中，至少一個胺基酸之該置換係在蛋白酶域之部分1中。在一實施例中，至少一個胺基酸之該置換係在蛋白酶域之部分2中。在一實施例中，該置換係選自由D130A、H193A、N295A及S512A組成之群。在一實施例中，該置換為D130A。在另一實施例中，該置換為H193A。在另一實施例中，該置換為N295A。在又一實施例中，該置換為S512A。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzyme-inactive fragment of SCPA, which includes a protease domain, a protease-associated domain (PA domain) and three fiber binding protein type III (Fn) domains but does not include an export signal pre-sequence, a post-sequence and a cell wall anchoring domain, wherein the inactivation is achieved by replacing at least one amino acid of the wild-type sequence. Preferably, the replacement of at least one amino acid is in the protease domain. In one embodiment, the replacement of at least one amino acid is in part 1 of the protease domain. In one embodiment, the replacement of at least one amino acid is in part 2 of the protease domain. In one embodiment, the replacement is selected from the group consisting of D130A, H193A, N295A and S512A. In one embodiment, the replacement is D130A. In another embodiment, the substitution is H193A. In another embodiment, the substitution is N295A. In yet another embodiment, the substitution is S512A.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCPB之片段，其包含蛋白酶域、蛋白酶相關域(PA域)及三個纖維結合蛋白III型(Fn)域但不包含輸出信號前序列、後序列及細胞壁錨定域，其中該不活化係藉由置換野生型序列之至少一個胺基酸來實現。較佳地，至少一個胺基酸之該置換係在蛋白酶域中。在一實施例中，至少一個胺基酸之該置換係在蛋白酶域之部分1中。在一實施例中，至少一個胺基酸之該置換係在蛋白酶域之部分2中。在一實施例中，該置換係選自由D130A、H193A、N295A及S512A組成之群。在一實施例中，該置換為D130A。在另一實施例中，該置換為H193A。在另一實施例中，該置換為N295A。在又一實施例中，該置換為S512A。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzyme-inactive fragment of SCPB, which includes a protease domain, a protease-associated domain (PA domain) and three fiber binding protein type III (Fn) domains but does not include an export signal pre-sequence, a post-sequence and a cell wall anchoring domain, wherein the inactivation is achieved by replacing at least one amino acid of the wild-type sequence. Preferably, the replacement of at least one amino acid is in the protease domain. In one embodiment, the replacement of at least one amino acid is in part 1 of the protease domain. In one embodiment, the replacement of at least one amino acid is in part 2 of the protease domain. In one embodiment, the replacement is selected from the group consisting of D130A, H193A, N295A and S512A. In one embodiment, the replacement is D130A. In another embodiment, the substitution is H193A. In another embodiment, the substitution is N295A. In yet another embodiment, the substitution is S512A.

在一實施例中，SCP之酶活性係藉由置換野生型序列之至少兩個胺基酸而不活化。在一實施例中，該至少兩個胺基酸置換係選自由D130A、H193A、N295A及S512A組成之群。在一實施例中，該至少兩個胺基酸置換為D130A及H193A。在一實施例中，該至少兩個胺基酸置換為D130A及N295A。在一實施例中，該至少兩個胺基酸置換為D130A及S512A。在一實施例中，該至少兩個胺基酸置換為H193A及N295A。在一實施例中，該至少兩個胺基酸置換為H193A及S512A。在一實施例中，該至少兩個胺基酸置換為N295A及S512A。In one embodiment, the enzymatic activity of SCP is inactivated by replacing at least two amino acids of the wild-type sequence. In one embodiment, the at least two amino acid replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In one embodiment, the at least two amino acid replacements are D130A and H193A. In one embodiment, the at least two amino acid replacements are D130A and N295A. In one embodiment, the at least two amino acid replacements are D130A and S512A. In one embodiment, the at least two amino acid replacements are H193A and N295A. In one embodiment, the at least two amino acid replacements are H193A and S512A. In one embodiment, the at least two amino acid substitutions are N295A and S512A.

因此，在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCP，其中該不活化係藉由置換野生型序列之至少兩個胺基酸來實現。較佳地，該至少兩個胺基酸置換係在蛋白酶域中。在一實施例中，該至少兩個胺基酸置換係在蛋白酶域之部分1中。在一實施例中，該至少兩個胺基酸置換係在蛋白酶域之部分2中。在一實施例中，該至少兩個胺基酸置換係選自由D130A、H193A、N295A及S512A組成之群。在一實施例中，該至少兩個胺基酸置換為D130A及H193A。在一實施例中，該至少兩個胺基酸置換為D130A及N295A。較佳地，該至少兩個胺基酸置換為D130A及S512A。在一實施例中，該至少兩個胺基酸置換為H193A及N295A。在一實施例中，該至少兩個胺基酸置換為H193A及S512A。在一實施例中，該至少兩個胺基酸置換為N295A及S512A。Therefore, in one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzyme-inactive SCP, wherein the inactivation is achieved by replacing at least two amino acids of the wild-type sequence. Preferably, the at least two amino acid replacements are in the protease domain. In one embodiment, the at least two amino acid replacements are in part 1 of the protease domain. In one embodiment, the at least two amino acid replacements are in part 2 of the protease domain. In one embodiment, the at least two amino acid replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In one embodiment, the at least two amino acid replacements are D130A and H193A. In one embodiment, the at least two amino acid replacements are D130A and N295A. Preferably, the at least two amino acids are replaced by D130A and S512A. In one embodiment, the at least two amino acids are replaced by H193A and N295A. In one embodiment, the at least two amino acids are replaced by H193A and S512A. In one embodiment, the at least two amino acids are replaced by N295A and S512A.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCPA，其中該不活化係藉由置換野生型序列之至少兩個胺基酸來實現。較佳地，該至少兩個胺基酸置換係在蛋白酶域中。在一實施例中，該至少兩個胺基酸置換係在蛋白酶域之部分1中。在一實施例中，該至少兩個胺基酸置換係在蛋白酶域之部分2中。在一實施例中，該至少兩個胺基酸置換係選自由D130A、H193A、N295A及S512A組成之群。在一實施例中，該至少兩個胺基酸置換為D130A及H193A。在一實施例中，該至少兩個胺基酸置換為D130A及N295A。較佳地，該至少兩個胺基酸置換為D130A及S512A。在一實施例中，該至少兩個胺基酸置換為H193A及N295A。在一實施例中，該至少兩個胺基酸置換為H193A及S512A。在一實施例中，該至少兩個胺基酸置換為N295A及S512A。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzyme-inactive SCPA, wherein the inactivation is achieved by replacing at least two amino acids of the wild-type sequence. Preferably, the at least two amino acid replacements are in the protease domain. In one embodiment, the at least two amino acid replacements are in part 1 of the protease domain. In one embodiment, the at least two amino acid replacements are in part 2 of the protease domain. In one embodiment, the at least two amino acid replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In one embodiment, the at least two amino acid replacements are D130A and H193A. In one embodiment, the at least two amino acid replacements are D130A and N295A. Preferably, the at least two amino acids are replaced by D130A and S512A. In one embodiment, the at least two amino acids are replaced by H193A and N295A. In one embodiment, the at least two amino acids are replaced by H193A and S512A. In one embodiment, the at least two amino acids are replaced by N295A and S512A.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCPB，其中該不活化係藉由置換野生型序列之至少兩個胺基酸來實現。較佳地，該至少兩個胺基酸置換係在蛋白酶域中。在一實施例中，該至少兩個胺基酸置換係在蛋白酶域之部分1中。在一實施例中，該至少兩個胺基酸置換係在蛋白酶域之部分2中。在一實施例中，該至少兩個胺基酸置換係選自由D130A、H193A、N295A及S512A組成之群。在一實施例中，該至少兩個胺基酸置換為D130A及H193A。在一實施例中，該至少兩個胺基酸置換為D130A及N295A。較佳地，該至少兩個胺基酸置換為D130A及S512A。在一實施例中，該至少兩個胺基酸置換為H193A及N295A。在一實施例中，該至少兩個胺基酸置換為H193A及S512A。在一實施例中，該至少兩個胺基酸置換為N295A及S512A。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzyme-inactive SCPB, wherein the inactivation is achieved by replacing at least two amino acids of the wild-type sequence. Preferably, the at least two amino acid replacements are in the protease domain. In one embodiment, the at least two amino acid replacements are in part 1 of the protease domain. In one embodiment, the at least two amino acid replacements are in part 2 of the protease domain. In one embodiment, the at least two amino acid replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In one embodiment, the at least two amino acid replacements are D130A and H193A. In one embodiment, the at least two amino acid replacements are D130A and N295A. Preferably, the at least two amino acids are replaced by D130A and S512A. In one embodiment, the at least two amino acids are replaced by H193A and N295A. In one embodiment, the at least two amino acids are replaced by H193A and S512A. In one embodiment, the at least two amino acids are replaced by N295A and S512A.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCP之片段，其中該不活化係藉由置換野生型序列之至少兩個胺基酸來實現。較佳地，該至少兩個胺基酸置換係在蛋白酶域中。在一實施例中，該至少兩個胺基酸置換係在蛋白酶域之部分1中。在一實施例中，該至少兩個胺基酸置換係在蛋白酶域之部分2中。在一實施例中，該至少兩個胺基酸置換係選自由D130A、H193A、N295A及S512A組成之群。在一實施例中，該至少兩個胺基酸置換為D130A及H193A。在一實施例中，該至少兩個胺基酸置換為D130A及N295A。較佳地，該至少兩個胺基酸置換為D130A及S512A。在一實施例中，該至少兩個胺基酸置換為H193A及N295A。在一實施例中，該至少兩個胺基酸置換為H193A及S512A。在一實施例中，該至少兩個胺基酸置換為N295A及S512A。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is a fragment of an enzyme-inactive SCP, wherein the inactivation is achieved by replacing at least two amino acids of the wild-type sequence. Preferably, the at least two amino acid replacements are in the protease domain. In one embodiment, the at least two amino acid replacements are in part 1 of the protease domain. In one embodiment, the at least two amino acid replacements are in part 2 of the protease domain. In one embodiment, the at least two amino acid replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In one embodiment, the at least two amino acid replacements are D130A and H193A. In one embodiment, the at least two amino acid replacements are D130A and N295A. Preferably, the at least two amino acids are replaced by D130A and S512A. In one embodiment, the at least two amino acids are replaced by H193A and N295A. In one embodiment, the at least two amino acids are replaced by H193A and S512A. In one embodiment, the at least two amino acids are replaced by N295A and S512A.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCP之片段，其包含蛋白酶域、蛋白酶相關域(PA域)及三個纖維結合蛋白III型(Fn)域但不包含輸出信號前序列、後序列及細胞壁錨定域，其中該不活化係藉由置換野生型序列之至少兩個胺基酸來實現。較佳地，該至少兩個胺基酸置換係在蛋白酶域中。在一實施例中，該至少兩個胺基酸置換係在蛋白酶域之部分1中。在一實施例中，該至少兩個胺基酸置換係在蛋白酶域之部分2中。在一實施例中，該至少兩個胺基酸置換係選自由D130A、H193A、N295A及S512A組成之群。在一實施例中，該至少兩個胺基酸置換為D130A及H193A。在一實施例中，該至少兩個胺基酸置換為D130A及N295A。較佳地，該至少兩個胺基酸置換為D130A及S512A。在一實施例中，該至少兩個胺基酸置換為H193A及N295A。在一實施例中，該至少兩個胺基酸置換為H193A及S512A。在一實施例中，該至少兩個胺基酸置換為N295A及S512A。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzyme-inactive fragment of SCP, which comprises a protease domain, a protease-associated domain (PA domain) and three fiber binding protein type III (Fn) domains but does not include an export signal pre-sequence, a post-sequence and a cell wall anchoring domain, wherein the inactivation is achieved by replacing at least two amino acids of the wild-type sequence. Preferably, the at least two amino acid substitutions are in the protease domain. In one embodiment, the at least two amino acid substitutions are in part 1 of the protease domain. In one embodiment, the at least two amino acid substitutions are in part 2 of the protease domain. In one embodiment, the at least two amino acid substitutions are selected from the group consisting of D130A, H193A, N295A and S512A. In one embodiment, the at least two amino acids are replaced by D130A and H193A. In one embodiment, the at least two amino acids are replaced by D130A and N295A. Preferably, the at least two amino acids are replaced by D130A and S512A. In one embodiment, the at least two amino acids are replaced by H193A and N295A. In one embodiment, the at least two amino acids are replaced by H193A and S512A. In one embodiment, the at least two amino acids are replaced by N295A and S512A.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCPA之片段，其包含蛋白酶域、蛋白酶相關域(PA域)及三個纖維結合蛋白III型(Fn)域但不包含輸出信號前序列、後序列及細胞壁錨定域，其中該不活化係藉由置換野生型序列之至少兩個胺基酸來實現。較佳地，該至少兩個胺基酸置換係在蛋白酶域中。在一實施例中，該至少兩個胺基酸置換係在蛋白酶域之部分1中。在一實施例中，該至少一個胺基酸置換係在蛋白酶域之部分2中。在一實施例中，該至少兩個胺基酸置換係選自由D130A、H193A、N295A及S512A組成之群。在一實施例中，該至少兩個胺基酸置換為D130A及H193A。在一實施例中，該至少兩個胺基酸置換為D130A及N295A。較佳地，該至少兩個胺基酸置換為D130A及S512A。在一實施例中，該至少兩個胺基酸置換為H193A及N295A。在一實施例中，該至少兩個胺基酸置換為H193A及S512A。在一實施例中，該至少兩個胺基酸置換為N295A及S512A。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzyme-inactive fragment of SCPA, which comprises a protease domain, a protease-associated domain (PA domain) and three fiber binding protein type III (Fn) domains but does not include an export signal pre-sequence, a post-sequence and a cell wall anchoring domain, wherein the inactivation is achieved by replacing at least two amino acids of the wild-type sequence. Preferably, the at least two amino acid substitutions are in the protease domain. In one embodiment, the at least two amino acid substitutions are in part 1 of the protease domain. In one embodiment, the at least one amino acid substitution is in part 2 of the protease domain. In one embodiment, the at least two amino acid substitutions are selected from the group consisting of D130A, H193A, N295A and S512A. In one embodiment, the at least two amino acids are replaced by D130A and H193A. In one embodiment, the at least two amino acids are replaced by D130A and N295A. Preferably, the at least two amino acids are replaced by D130A and S512A. In one embodiment, the at least two amino acids are replaced by H193A and N295A. In one embodiment, the at least two amino acids are replaced by H193A and S512A. In one embodiment, the at least two amino acids are replaced by N295A and S512A.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCPB之片段，其包含蛋白酶域、蛋白酶相關域(PA域)及三個纖維結合蛋白III型(Fn)域但不包含輸出信號前序列、後序列及細胞壁錨定域，其中該不活化係藉由置換野生型序列之至少兩個胺基酸來實現。較佳地，該至少兩個胺基酸置換係在蛋白酶域中。在一實施例中，該至少兩個胺基酸置換係在蛋白酶域之部分1中。在一實施例中，該至少兩個胺基酸置換係在蛋白酶域之部分2中。在一實施例中，該至少兩個胺基酸置換係選自由D130A、H193A、N295A及S512A組成之群。在一實施例中，該至少兩個胺基酸置換為D130A及H193A。在一實施例中，該至少兩個胺基酸置換為D130A及N295A。較佳地，該至少兩個胺基酸置換為D130A及S512A。在一實施例中，該至少兩個胺基酸置換為H193A及N295A。在一實施例中，該至少兩個胺基酸置換為H193A及S512A。在一實施例中，該至少兩個胺基酸置換為N295A及S512A。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzyme-inactive fragment of SCPB, which comprises a protease domain, a protease-associated domain (PA domain) and three fiber binding protein type III (Fn) domains but does not comprise an export signal pre-sequence, a post-sequence and a cell wall anchoring domain, wherein the inactivation is achieved by replacing at least two amino acids of the wild-type sequence. Preferably, the at least two amino acid substitutions are in the protease domain. In one embodiment, the at least two amino acid substitutions are in part 1 of the protease domain. In one embodiment, the at least two amino acid substitutions are in part 2 of the protease domain. In one embodiment, the at least two amino acid substitutions are selected from the group consisting of D130A, H193A, N295A and S512A. In one embodiment, the at least two amino acids are replaced by D130A and H193A. In one embodiment, the at least two amino acids are replaced by D130A and N295A. Preferably, the at least two amino acids are replaced by D130A and S512A. In one embodiment, the at least two amino acids are replaced by H193A and N295A. In one embodiment, the at least two amino acids are replaced by H193A and S512A. In one embodiment, the at least two amino acids are replaced by N295A and S512A.

在一實施例中，SCP之酶活性係藉由置換野生型序列之至少三個胺基酸而不活化。在一實施例中，該至少三個胺基酸置換係選自由D130A、H193A、N295A及S512A組成之群。在一實施例中，該至少三個胺基酸置換為D130A、H193A及N295A。在一實施例中，該至少三個胺基酸置換為D130A、H193A及S512A。在一實施例中，該至少三個胺基酸置換為D130A、N295A及S512A。在一實施例中，該至少三個胺基酸置換為H193A、N295A及S512A。In one embodiment, the enzymatic activity of SCP is inactivated by replacing at least three amino acids of the wild-type sequence. In one embodiment, the at least three amino acid replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In one embodiment, the at least three amino acid replacements are D130A, H193A and N295A. In one embodiment, the at least three amino acid replacements are D130A, H193A and S512A. In one embodiment, the at least three amino acid replacements are D130A, N295A and S512A. In one embodiment, the at least three amino acid replacements are H193A, N295A and S512A.

因此，在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCP，其中該不活化係藉由置換野生型序列之至少三個胺基酸來實現。較佳地，該至少三個胺基酸置換係在蛋白酶域中。在一實施例中，該至少三個胺基酸置換係在蛋白酶域之部分1中。在一實施例中，該至少三個胺基酸置換係在蛋白酶域之部分2中。在一實施例中，該至少三個胺基酸置換係選自由D130A、H193A、N295A及S512A組成之群。在一實施例中，該至少三個胺基酸置換為D130A、H193A及N295A。在一實施例中，該至少三個胺基酸置換為D130A、H193A及S512A。在一實施例中，該至少三個胺基酸置換為D130A、N295A及S512A。在一實施例中，該至少三個胺基酸置換為H193A、N295A及S512A。Therefore, in one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzyme-inactive SCP, wherein the inactivation is achieved by replacing at least three amino acids of the wild-type sequence. Preferably, the at least three amino acid replacements are in the protease domain. In one embodiment, the at least three amino acid replacements are in part 1 of the protease domain. In one embodiment, the at least three amino acid replacements are in part 2 of the protease domain. In one embodiment, the at least three amino acid replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In one embodiment, the at least three amino acid replacements are D130A, H193A and N295A. In one embodiment, the at least three amino acid replacements are D130A, H193A and S512A. In one embodiment, the at least three amino acids are replaced by D130A, N295A and S512A. In one embodiment, the at least three amino acids are replaced by H193A, N295A and S512A.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCPA，其中該不活化係藉由置換野生型序列之至少三個胺基酸來實現。較佳地，該至少三個胺基酸置換係在蛋白酶域中。在一實施例中，該至少三個胺基酸置換係在蛋白酶域之部分1中。在一實施例中，該至少三個胺基酸置換係在蛋白酶域之部分2中。在一實施例中，該至少三個胺基酸置換係選自由D130A、H193A、N295A及S512A組成之群。在一實施例中，該至少三個胺基酸置換為D130A、H193A及N295A。在一實施例中，該至少三個胺基酸置換為D130A、H193A及S512A。在一實施例中，該至少三個胺基酸置換為D130A、N295A及S512A。在一實施例中，該至少三個胺基酸置換為H193A、N295A及S512A。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzyme-inactive SCPA, wherein the inactivation is achieved by replacing at least three amino acids of the wild-type sequence. Preferably, the at least three amino acid replacements are in the protease domain. In one embodiment, the at least three amino acid replacements are in part 1 of the protease domain. In one embodiment, the at least three amino acid replacements are in part 2 of the protease domain. In one embodiment, the at least three amino acid replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In one embodiment, the at least three amino acid replacements are D130A, H193A and N295A. In one embodiment, the at least three amino acid replacements are D130A, H193A and S512A. In one embodiment, the at least three amino acids are replaced by D130A, N295A and S512A. In one embodiment, the at least three amino acids are replaced by H193A, N295A and S512A.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCPB，其中該不活化係藉由置換野生型序列之至少三個胺基酸來實現。較佳地，該至少三個胺基酸置換係在蛋白酶域中。在一實施例中，該至少三個胺基酸置換係在蛋白酶域之部分1中。在一實施例中，該至少三個胺基酸置換係在蛋白酶域之部分2中。在一實施例中，該至少三個胺基酸置換係選自由D130A、H193A、N295A及S512A組成之群。在一實施例中，該至少三個胺基酸置換為D130A、H193A及N295A。在一實施例中，該至少三個胺基酸置換為D130A、H193A及S512A。在一實施例中，該至少三個胺基酸置換為D130A、N295A及S512A。在一實施例中，該至少三個胺基酸置換為H193A、N295A及S512A。In one embodiment, the carrier protein of the serotype 38 carbohydrate conjugate of the present invention is an enzyme-inactivated SCPB, wherein the inactivation is achieved by replacing at least three amino acids of the wild-type sequence. Preferably, the at least three amino acid replacements are in the protease domain. In one embodiment, the at least three amino acid replacements are in part 1 of the protease domain. In one embodiment, the at least three amino acid replacements are in part 2 of the protease domain. In one embodiment, the at least three amino acid replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In one embodiment, the at least three amino acid replacements are D130A, H193A and N295A. In one embodiment, the at least three amino acid replacements are D130A, H193A and S512A. In one embodiment, the at least three amino acids are replaced by D130A, N295A and S512A. In one embodiment, the at least three amino acids are replaced by H193A, N295A and S512A.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCP之片段，其中該不活化係藉由置換野生型序列之至少三個胺基酸來實現。較佳地，該至少三個胺基酸置換係在蛋白酶域中。在一實施例中，該至少三個胺基酸置換係在蛋白酶域之部分1中。在一實施例中，該至少三個胺基酸置換係在蛋白酶域之部分2中。在一實施例中，該至少三個胺基酸置換係選自由D130A、H193A、N295A及S512A組成之群。在一實施例中，該至少三個胺基酸置換為D130A、H193A及N295A。在一實施例中，該至少三個胺基酸置換為D130A、H193A及S512A。在一實施例中，該至少三個胺基酸置換為D130A、N295A及S512A。在一實施例中，該至少三個胺基酸置換為H193A、N295A及S512A。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is a fragment of an enzyme-inactive SCP, wherein the inactivation is achieved by replacing at least three amino acids of the wild-type sequence. Preferably, the at least three amino acid replacements are in the protease domain. In one embodiment, the at least three amino acid replacements are in part 1 of the protease domain. In one embodiment, the at least three amino acid replacements are in part 2 of the protease domain. In one embodiment, the at least three amino acid replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In one embodiment, the at least three amino acid replacements are D130A, H193A and N295A. In one embodiment, the at least three amino acid replacements are D130A, H193A and S512A. In one embodiment, the at least three amino acids are replaced by D130A, N295A and S512A. In one embodiment, the at least three amino acids are replaced by H193A, N295A and S512A.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCP之片段，其包含蛋白酶域、蛋白酶相關域(PA域)及三個纖維結合蛋白III型(Fn)域但不包含輸出信號前序列、後序列及細胞壁錨定域，其中該不活化係藉由置換野生型序列之至少三個胺基酸來實現。較佳地，該至少三個胺基酸置換係在蛋白酶域中。在一實施例中，該至少三個胺基酸置換係在蛋白酶域之部分1中。在一實施例中，該至少三個胺基酸置換係在蛋白酶域之部分2中。在一實施例中，該至少三個胺基酸置換係選自由D130A、H193A、N295A及S512A組成之群。在一實施例中，該至少三個胺基酸置換為D130A、H193A及N295A。在一實施例中，該至少三個胺基酸置換為D130A、H193A及S512A。在一實施例中，該至少三個胺基酸置換為D130A、N295A及S512A。在一實施例中，該至少三個胺基酸置換為H193A、N295A及S512A。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzyme-inactive fragment of SCP, which comprises a protease domain, a protease-associated domain (PA domain) and three fiber binding protein type III (Fn) domains but does not comprise an export signal pre-sequence, a post-sequence and a cell wall anchoring domain, wherein the inactivation is achieved by replacing at least three amino acids of the wild-type sequence. Preferably, the at least three amino acid substitutions are in the protease domain. In one embodiment, the at least three amino acid substitutions are in part 1 of the protease domain. In one embodiment, the at least three amino acid substitutions are in part 2 of the protease domain. In one embodiment, the at least three amino acid substitutions are selected from the group consisting of D130A, H193A, N295A and S512A. In one embodiment, the at least three amino acid replacements are D130A, H193A and N295A. In one embodiment, the at least three amino acid replacements are D130A, H193A and S512A. In one embodiment, the at least three amino acid replacements are D130A, N295A and S512A. In one embodiment, the at least three amino acid replacements are H193A, N295A and S512A.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCPA之片段，其包含蛋白酶域、蛋白酶相關域(PA域)及三個纖維結合蛋白III型(Fn)域但不包含輸出信號前序列、後序列及細胞壁錨定域，其中該不活化係藉由置換野生型序列之至少三個胺基酸來實現。較佳地，該至少三個胺基酸置換係在蛋白酶域中。在一實施例中，該至少三個胺基酸置換係在蛋白酶域之部分1中。在一實施例中，該至少三個胺基酸置換係在蛋白酶域之部分2中。在一實施例中，該至少三個胺基酸置換係選自由D130A、H193A、N295A及S512A組成之群。在一實施例中，該至少三個胺基酸置換為D130A、H193A及N295A。在一實施例中，該至少三個胺基酸置換為D130A、H193A及S512A。在一實施例中，該至少三個胺基酸置換為D130A、N295A及S512A。在一實施例中，該至少三個胺基酸置換為H193A、N295A及S512A。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzyme-inactive fragment of SCPA, which comprises a protease domain, a protease-associated domain (PA domain) and three fiber binding protein type III (Fn) domains but does not comprise an export signal pre-sequence, a post-sequence and a cell wall anchoring domain, wherein the inactivation is achieved by replacing at least three amino acids of the wild-type sequence. Preferably, the at least three amino acid substitutions are in the protease domain. In one embodiment, the at least three amino acid substitutions are in part 1 of the protease domain. In one embodiment, the at least three amino acid substitutions are in part 2 of the protease domain. In one embodiment, the at least three amino acid substitutions are selected from the group consisting of D130A, H193A, N295A and S512A. In one embodiment, the at least three amino acid replacements are D130A, H193A and N295A. In one embodiment, the at least three amino acid replacements are D130A, H193A and S512A. In one embodiment, the at least three amino acid replacements are D130A, N295A and S512A. In one embodiment, the at least three amino acid replacements are H193A, N295A and S512A.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCPB之片段，其包含蛋白酶域、蛋白酶相關域(PA域)及三個纖維結合蛋白III型(Fn)域但不包含輸出信號前序列、後序列及細胞壁錨定域，其中該不活化係藉由置換野生型序列之至少三個胺基酸來實現。較佳地，該至少三個胺基酸置換係在蛋白酶域中。在一實施例中，該至少三個胺基酸置換係在蛋白酶域之部分1中。在一實施例中，該至少三個胺基酸置換係在蛋白酶域之部分2中。在一實施例中，該至少三個胺基酸置換係選自由D130A、H193A、N295A及S512A組成之群。在一實施例中，該至少三個胺基酸置換為D130A、H193A及N295A。在一實施例中，該至少三個胺基酸置換為D130A、H193A及S512A。在一實施例中，該至少三個胺基酸置換為D130A、N295A及S512A。在一實施例中，該至少三個胺基酸置換為H193A、N295A及S512A。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzyme-inactive fragment of SCPB, which comprises a protease domain, a protease-associated domain (PA domain) and three fiber binding protein type III (Fn) domains but does not comprise an export signal pre-sequence, a post-sequence and a cell wall anchoring domain, wherein the inactivation is achieved by replacing at least three amino acids of the wild-type sequence. Preferably, the at least three amino acid substitutions are in the protease domain. In one embodiment, the at least three amino acid substitutions are in part 1 of the protease domain. In one embodiment, the at least three amino acid substitutions are in part 2 of the protease domain. In one embodiment, the at least three amino acid substitutions are selected from the group consisting of D130A, H193A, N295A and S512A. In one embodiment, the at least three amino acid replacements are D130A, H193A and N295A. In one embodiment, the at least three amino acid replacements are D130A, H193A and S512A. In one embodiment, the at least three amino acid replacements are D130A, N295A and S512A. In one embodiment, the at least three amino acid replacements are H193A, N295A and S512A.

在一實施例中，SCP之酶活性係藉由置換野生型序列之至少四個胺基酸而不活化。在一實施例中，該至少四個胺基酸置換為D130A、H193A、N295A及S512A。In one embodiment, the enzymatic activity of SCP is inactivated by replacing at least four amino acids of the wild-type sequence. In one embodiment, the at least four amino acids are replaced by D130A, H193A, N295A and S512A.

因此，在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCP，其中該不活化係藉由置換野生型序列之至少四個胺基酸來實現。較佳地，該至少四個胺基酸置換係在蛋白酶域中。在一實施例中，該至少四個胺基酸置換係在蛋白酶域之部分1中。在一實施例中，該至少四個胺基酸置換係在蛋白酶域之部分2中。在一實施例中，該至少四個胺基酸置換為D130A、H193A、N295A及S512A。Thus, in one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzymatically inactive SCP, wherein the inactivation is achieved by replacing at least four amino acids of the wild-type sequence. Preferably, the at least four amino acid substitutions are in the protease domain. In one embodiment, the at least four amino acid substitutions are in part 1 of the protease domain. In one embodiment, the at least four amino acid substitutions are in part 2 of the protease domain. In one embodiment, the at least four amino acid substitutions are D130A, H193A, N295A and S512A.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCPA，其中該不活化係藉由置換野生型序列之至少四個胺基酸來實現。較佳地，該至少四個胺基酸置換係在蛋白酶域中。在一實施例中，該至少四個胺基酸置換係在蛋白酶域之部分1中。在一實施例中，該至少四個胺基酸置換係在蛋白酶域之部分2中。在一實施例中，該至少四個胺基酸置換為D130A、H193A、N295A及S512A。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzymatically inactive SCPA, wherein the inactivation is achieved by replacing at least four amino acids of the wild-type sequence. Preferably, the at least four amino acid substitutions are in the protease domain. In one embodiment, the at least four amino acid substitutions are in part 1 of the protease domain. In one embodiment, the at least four amino acid substitutions are in part 2 of the protease domain. In one embodiment, the at least four amino acid substitutions are D130A, H193A, N295A and S512A.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCPB，其中該不活化係藉由置換野生型序列之至少四個胺基酸來實現。較佳地，該至少四個胺基酸置換係在蛋白酶域中。在一實施例中，該至少四個胺基酸置換係在蛋白酶域之部分1中。在一實施例中，該至少四個胺基酸置換係在蛋白酶域之部分2中。在一實施例中，該至少四個胺基酸置換為D130A、H193A、N295A及S512A。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzymatically inactive SCPB, wherein the inactivation is achieved by replacing at least four amino acids of the wild-type sequence. Preferably, the at least four amino acid substitutions are in the protease domain. In one embodiment, the at least four amino acid substitutions are in part 1 of the protease domain. In one embodiment, the at least four amino acid substitutions are in part 2 of the protease domain. In one embodiment, the at least four amino acid substitutions are D130A, H193A, N295A and S512A.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCP之片段，其中該不活化係藉由置換野生型序列之至少四個胺基酸來實現。較佳地，該至少四個胺基酸置換係在蛋白酶域中。在一實施例中，該至少四個胺基酸置換係在蛋白酶域之部分1中。在一實施例中，該至少四個胺基酸置換係在蛋白酶域之部分2中。在一實施例中，該至少四個胺基酸置換為D130A、H193A、N295A及S512A。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzymatically inactive fragment of SCP, wherein the inactivation is achieved by replacing at least four amino acids of the wild-type sequence. Preferably, the at least four amino acid substitutions are in the protease domain. In one embodiment, the at least four amino acid substitutions are in part 1 of the protease domain. In one embodiment, the at least four amino acid substitutions are in part 2 of the protease domain. In one embodiment, the at least four amino acid substitutions are D130A, H193A, N295A and S512A.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCP之片段，其包含蛋白酶域、蛋白酶相關域(PA域)及三個纖維結合蛋白III型(Fn)域但不包含輸出信號前序列、後序列及細胞壁錨定域，其中該不活化係藉由置換野生型序列之至少四個胺基酸來實現。較佳地，該至少四個胺基酸置換係在蛋白酶域中。在一實施例中，該至少四個胺基酸置換係在蛋白酶域之部分1中。在一實施例中，該至少四個胺基酸置換係在蛋白酶域之部分2中。在一實施例中，該至少四個胺基酸置換為D130A、H193A、N295A及S512A。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzyme-inactive fragment of SCP, which comprises a protease domain, a protease-associated domain (PA domain) and three fiber binding protein type III (Fn) domains but does not comprise an export signal pre-sequence, a post-sequence and a cell wall anchoring domain, wherein the inactivation is achieved by replacing at least four amino acids of the wild-type sequence. Preferably, the at least four amino acid substitutions are in the protease domain. In one embodiment, the at least four amino acid substitutions are in part 1 of the protease domain. In one embodiment, the at least four amino acid substitutions are in part 2 of the protease domain. In one embodiment, the at least four amino acid substitutions are D130A, H193A, N295A and S512A.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCPA之片段，其包含蛋白酶域、蛋白酶相關域(PA域)及三個纖維結合蛋白III型(Fn)域但不包含輸出信號前序列、後序列及細胞壁錨定域，其中該不活化係藉由置換野生型序列之至少四個胺基酸來實現。較佳地，該至少四個胺基酸置換係在蛋白酶域中。在一實施例中，該至少四個胺基酸置換係在蛋白酶域之部分1中。在一實施例中，該至少一個胺基酸置換係在蛋白酶域之部分2中。在一實施例中，該至少四個胺基酸置換為D130A、H193A、N295A及S512A。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzyme-inactive fragment of SCPA, which comprises a protease domain, a protease-associated domain (PA domain) and three fiber binding protein type III (Fn) domains but does not include an export signal pre-sequence, a post-sequence and a cell wall anchoring domain, wherein the inactivation is achieved by replacing at least four amino acids of the wild-type sequence. Preferably, the at least four amino acid substitutions are in the protease domain. In one embodiment, the at least four amino acid substitutions are in part 1 of the protease domain. In one embodiment, the at least one amino acid substitution is in part 2 of the protease domain. In one embodiment, the at least four amino acid substitutions are D130A, H193A, N295A and S512A.

在一實施例中，本發明之血清型38醣結合物之載體蛋白為酶不活化的SCPB之片段，其包含蛋白酶域、蛋白酶相關域(PA域)及三個纖維結合蛋白III型(Fn)域但不包含輸出信號前序列、後序列及細胞壁錨定域，其中該不活化係藉由置換野生型序列之至少四個胺基酸來實現。較佳地，該至少四個胺基酸置換係在蛋白酶域中。在一實施例中，該至少四個胺基酸置換係在蛋白酶域之部分1中。在一實施例中，該至少四個胺基酸置換係在蛋白酶域之部分2中。在一實施例中，該至少四個胺基酸置換為D130A、H193A、N295A及S512A。In one embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzyme-inactive fragment of SCPB, which includes a protease domain, a protease-associated domain (PA domain) and three fiber binding protein type III (Fn) domains but does not include an export signal pre-sequence, a post-sequence and a cell wall anchoring domain, wherein the inactivation is achieved by replacing at least four amino acids of the wild-type sequence. Preferably, the at least four amino acid substitutions are in the protease domain. In one embodiment, the at least four amino acid substitutions are in part 1 of the protease domain. In one embodiment, the at least four amino acid substitutions are in part 2 of the protease domain. In one embodiment, the at least four amino acid substitutions are D130A, H193A, N295A and S512A.

在一特定實施例中，本發明之血清型38醣結合物之載體蛋白係由SEQ ID NO: 1組成的酶不活化的SCP之片段。In a specific embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzymatically inactive fragment of SCP consisting of SEQ ID NO: 1.

在一特定實施例中，本發明之血清型38醣結合物之載體蛋白係由SEQ ID NO: 2組成的酶不活化的SCP之片段。In a specific embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzymatically inactive fragment of SCP consisting of SEQ ID NO: 2.

SEQ ID NO: 1： SEQ ID NO: 1:

SEQ ID NO: 1之長度為950個胺基酸。The length of SEQ ID NO: 1 is 950 amino acids.

SEQ ID NO: 2： SEQ ID NO: 2:

SEQ ID NO: 2之長度為949個胺基酸。The length of SEQ ID NO: 2 is 949 amino acids.

在一特定實施例中，本發明之血清型38醣結合物之載體蛋白係由與SEQ ID NO: 1具有至少90%一致性之多肽組成的酶不活化的SCP之片段。In a specific embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 90% identity to SEQ ID NO: 1.

在一特定實施例中，本發明之血清型38醣結合物之載體蛋白係由與SEQ ID NO: 1具有至少95%一致性之多肽組成的酶不活化的SCP之片段。In a specific embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 95% identity to SEQ ID NO: 1.

在一特定實施例中，本發明之血清型38醣結合物之載體蛋白係由與SEQ ID NO: 1具有至少99%一致性之多肽組成的酶不活化的SCP之片段。In a specific embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 99% identity to SEQ ID NO: 1.

在一特定實施例中，本發明之血清型38醣結合物之載體蛋白係由與SEQ ID NO: 1具有至少99.5%一致性之多肽組成的酶不活化的SCP之片段。In a specific embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 99.5% identity to SEQ ID NO: 1.

在一特定實施例中，本發明之血清型38醣結合物之載體蛋白係由與SEQ ID NO: 1具有至少99.8%一致性之多肽組成的酶不活化的SCP之片段。In a specific embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 99.8% identity to SEQ ID NO: 1.

在一特定實施例中，本發明之血清型38醣結合物之載體蛋白係由與SEQ ID NO: 1具有至少99.85%一致性之多肽組成的酶不活化的SCP之片段。In a specific embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 99.85% identity to SEQ ID NO: 1.

在一特定實施例中，本發明之血清型38醣結合物之載體蛋白係由與SEQ ID NO: 2具有至少90%一致性之多肽組成的酶不活化的SCP之片段。In a specific embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 90% identity to SEQ ID NO: 2.

在一特定實施例中，本發明之血清型38醣結合物之載體蛋白係由與SEQ ID NO: 2具有至少95%一致性之多肽組成的酶不活化的SCP之片段。In a specific embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 95% identity to SEQ ID NO: 2.

在一特定實施例中，本發明之血清型38醣結合物之載體蛋白係由與SEQ ID NO: 2具有至少99%一致性之多肽組成的酶不活化的SCP之片段。In a specific embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 99% identity to SEQ ID NO: 2.

在一特定實施例中，本發明之血清型38醣結合物之載體蛋白係由與SEQ ID NO: 2具有至少99.5%一致性之多肽組成的酶不活化的SCP之片段。In a specific embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 99.5% identity to SEQ ID NO: 2.

在一特定實施例中，本發明之血清型38醣結合物之載體蛋白係由與SEQ ID NO: 2具有至少99.8%一致性之多肽組成的酶不活化的SCP之片段。In a specific embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 99.8% identity to SEQ ID NO: 2.

在一特定實施例中，本發明之血清型38醣結合物之載體蛋白係由與SEQ ID NO: 2具有至少99.85%一致性之多肽組成的酶不活化的SCP之片段。In a specific embodiment, the carrier protein of the serotype 38 glycoconjugate of the present invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 99.85% identity to SEQ ID NO: 2.

3. 免疫原組合物在一實施例中，本發明係關於一種包含本發明之肺炎鏈球菌血清型38醣之免疫原組合物。 3. Immunogenic Compositions In one embodiment, the present invention relates to an immunogenic composition comprising the S. pneumoniae serotype 38 saccharide of the present invention.

在一實施例中，本發明係關於一種包含本發明之肺炎鏈球菌血清型38醣結合物之免疫原組合物。In one embodiment, the present invention relates to an immunogenic composition comprising the S. pneumoniae serotype 38 saccharide conjugate of the present invention.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且包含1至45種不同醣結合物。In one embodiment, the present invention relates to an immunogenic composition comprising the S. pneumoniae serotype 38 saccharide conjugate of the present invention and comprising 1 to 45 different saccharide conjugates.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且包含1至45種來自不同肺炎鏈球菌血清型之醣結合物(1至45種肺炎鏈球菌結合物)。In one embodiment, the present invention relates to an immunogenic composition comprising the pneumococcal serotype 38 saccharide conjugate of the present invention and 1 to 45 saccharide conjugates from different pneumococcal serotypes (1 to 45 pneumococcal conjugates).

在一個實施例中，本發明係關於一種免疫原組合物，其包含來自7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24或25種不同肺炎鏈球菌血清型之醣結合物。在一個實施例中，免疫原組合物包含來自16或20種不同肺炎鏈球菌血清型之醣結合物。In one embodiment, the invention relates to an immunogenic composition comprising saccharide conjugates from 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 different serotypes of Streptococcus pneumoniae. In one embodiment, the immunogenic composition comprises saccharide conjugates from 16 or 20 different serotypes of Streptococcus pneumoniae.

在一實施例中，免疫原組合物為21、22、23、24或25價肺炎鏈球菌結合物組合物。In one embodiment, the immunogenic composition is a 21-, 22-, 23-, 24-, or 25-valent Streptococcus pneumoniae conjugate composition.

在一實施例中，免疫原組合物為21價肺炎鏈球菌結合物組合物。在一實施例中，免疫原組合物為22價肺炎鏈球菌結合物組合物。在一實施例中，免疫原組合物為23價肺炎鏈球菌結合物組合物。在一實施例中，免疫原組合物為24價肺炎鏈球菌結合物組合物。在一實施例中，免疫原組合物為25價肺炎鏈球菌結合物組合物。In one embodiment, the immunogenic composition is a 21-valent pneumococcal conjugate composition. In one embodiment, the immunogenic composition is a 22-valent pneumococcal conjugate composition. In one embodiment, the immunogenic composition is a 23-valent pneumococcal conjugate composition. In one embodiment, the immunogenic composition is a 24-valent pneumococcal conjugate composition. In one embodiment, the immunogenic composition is a 25-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且包含26至45種來自不同肺炎鏈球菌血清型之醣結合物(26至45種肺炎鏈球菌結合物)。在一個實施例中，本發明係關於一種免疫原組合物，其包含來自26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、41、42、43、44或45種不同肺炎鏈球菌血清型之醣結合物。In one embodiment, the present invention relates to an immunogenic composition comprising the pneumococcal serotype 38 saccharide conjugate of the present invention and 26 to 45 saccharide conjugates from different pneumococcal serotypes (26 to 45 pneumococcal conjugates). In one embodiment, the present invention relates to an immunogenic composition comprising saccharide conjugates from 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44 or 45 different pneumococcal serotypes.

在一個實施例中，免疫原組合物包含來自35或45種不同肺炎鏈球菌血清型之醣結合物。在一實施例中，免疫原組合物為35、36、37、38、39、40、41、42、43、44或45價肺炎鏈球菌結合物組合物。在一實施例中，免疫原組合物為40、41、42、43、44或45價肺炎鏈球菌結合物組合物。在一實施例中，免疫原組合物為40價肺炎鏈球菌結合物組合物。在一實施例中，免疫原組合物為41價肺炎鏈球菌結合物組合物。在一實施例中，免疫原組合物為42價肺炎鏈球菌結合物組合物。在一實施例中，免疫原組合物為43價肺炎鏈球菌結合物組合物。在一實施例中，免疫原組合物為44價肺炎鏈球菌結合物組合物。在一實施例中，免疫原組合物為45價肺炎鏈球菌結合物組合物。In one embodiment, the immunogenic composition comprises saccharide conjugates from 35 or 45 different pneumococcal serotypes. In one embodiment, the immunogenic composition is a 35, 36, 37, 38, 39, 40, 41, 42, 43, 44 or 45 valent pneumococcal conjugate composition. In one embodiment, the immunogenic composition is a 40, 41, 42, 43, 44 or 45 valent pneumococcal conjugate composition. In one embodiment, the immunogenic composition is a 40 valent pneumococcal conjugate composition. In one embodiment, the immunogenic composition is a 41 valent pneumococcal conjugate composition. In one embodiment, the immunogenic composition is a 42 valent pneumococcal conjugate composition. In one embodiment, the immunogenic composition is a 43 valent pneumococcal conjugate composition. In one embodiment, the immunogenic composition is a 44-valent pneumococcal conjugate composition. In one embodiment, the immunogenic composition is a 45-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含來自肺炎鏈球菌血清型4、6B、9V、14、18C、19F及23F之醣結合物。In one embodiment, the present invention relates to an immunogenic composition comprising the S. pneumoniae serotype 38 saccharide conjugate of the present invention and further comprising saccharide conjugates from S. pneumoniae serotypes 4, 6B, 9V, 14, 18C, 19F and 23F.

在一實施例中，該免疫原組合物另外包含來自肺炎鏈球菌血清型1、5及7F之醣結合物。In one embodiment, the immunogenic composition further comprises saccharide conjugates from S. pneumoniae serotypes 1, 5, and 7F.

在一實施例中，以上免疫原組合物中之任一者另外包含來自肺炎鏈球菌血清型3之醣結合物。In one embodiment, any of the above immunogenic compositions further comprises a saccharide conjugate from Streptococcus pneumoniae serotype 3.

在一實施例中，以上免疫原組合物中之任一者另外包含來自肺炎鏈球菌血清型6A及19A之醣結合物。In one embodiment, any of the above immunogenic compositions further comprises saccharide conjugates from Streptococcus pneumoniae serotypes 6A and 19A.

在一實施例中，以上免疫原組合物中之任一者另外包含來自肺炎鏈球菌血清型22F及33F之醣結合物。In one embodiment, any of the above immunogenic compositions further comprises saccharide conjugates from Streptococcus pneumoniae serotypes 22F and 33F.

在一實施例中，以上免疫原組合物中之任一者另外包含來自肺炎鏈球菌血清型8、10A、11A、12F及15B之醣結合物。In one embodiment, any of the above immunogenic compositions further comprises a saccharide conjugate from Streptococcus pneumoniae serotypes 8, 10A, 11A, 12F and 15B.

在一實施例中，以上免疫原組合物中之任一者另外包含來自肺炎鏈球菌血清型2之醣結合物。In one embodiment, any of the above immunogenic compositions further comprises a saccharide conjugate from Streptococcus pneumoniae serotype 2.

在一實施例中，以上免疫原組合物中之任一者另外包含來自肺炎鏈球菌血清型9N之醣結合物。In one embodiment, any of the above immunogenic compositions further comprises a saccharide conjugate from Streptococcus pneumoniae serotype 9N.

在一實施例中，以上免疫原組合物中之任一者另外包含來自肺炎鏈球菌血清型17F之醣結合物。In one embodiment, any of the above immunogenic compositions further comprises a saccharide conjugate from Streptococcus pneumoniae serotype 17F.

在一實施例中，以上免疫原組合物中之任一者另外包含來自肺炎鏈球菌血清型20之醣結合物。In one embodiment, any of the above immunogenic compositions further comprises a saccharide conjugate from Streptococcus pneumoniae serotype 20.

在一實施例中，以上免疫原組合物中之任一者另外包含來自肺炎鏈球菌血清型15C之醣結合物。In one embodiment, any of the above immunogenic compositions further comprises a saccharide conjugate from Streptococcus pneumoniae serotype 15C.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含來自肺炎鏈球菌血清型4、6B、9V、14、18C、19F及23F之醣結合物。在一實施例中，免疫原組合物為8價肺炎鏈球菌結合物組合物。In one embodiment, the present invention relates to an immunogenic composition, which comprises the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprises saccharide conjugates from pneumococcal serotypes 4, 6B, 9V, 14, 18C, 19F and 23F. In one embodiment, the immunogenic composition is an 8-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含來自肺炎鏈球菌血清型1、4、5、6B、7F、9V、14、18C、19F及23F之醣結合物。在一實施例中，免疫原組合物為11價肺炎鏈球菌結合物組合物。In one embodiment, the present invention relates to an immunogenic composition, which comprises the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprises saccharide conjugates from pneumococcal serotypes 1, 4, 5, 6B, 7F, 9V, 14, 18C, 19F and 23F. In one embodiment, the immunogenic composition is an 11-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含來自肺炎鏈球菌血清型1、3、4、5、6A、6B、7F、9V、14、18C、19A、19F及23F之醣結合物。在一實施例中，免疫原組合物為14價肺炎鏈球菌結合物組合物。In one embodiment, the present invention relates to an immunogenic composition comprising the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprising saccharide conjugates from pneumococcal serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F and 23F. In one embodiment, the immunogenic composition is a 14-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含來自肺炎鏈球菌血清型1、3、4、5、6A、6B、7F、9V、14、18C、19A、19F、22F、23F及33F之醣結合物。在一實施例中，免疫原組合物為16價肺炎鏈球菌結合物組合物。In one embodiment, the present invention relates to an immunogenic composition comprising the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprising saccharide conjugates from pneumococcal serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, 22F, 23F and 33F. In one embodiment, the immunogenic composition is a 16-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含來自肺炎鏈球菌血清型1、3、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15B、18C、19A、19F、22F、23F及33F之醣結合物。在一實施例中，免疫原組合物為21價肺炎鏈球菌結合物組合物。In one embodiment, the present invention relates to an immunogenic composition comprising the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprising saccharide conjugates from pneumococcal serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F and 33F. In one embodiment, the immunogenic composition is a 21-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含來自肺炎鏈球菌血清型1、3、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15C、18C、19A、19F、22F、23F及33F之醣結合物。在一實施例中，免疫原組合物為21價肺炎鏈球菌結合物組合物。In one embodiment, the present invention relates to an immunogenic composition comprising the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprising saccharide conjugates from pneumococcal serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15C, 18C, 19A, 19F, 22F, 23F and 33F. In one embodiment, the immunogenic composition is a 21-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含來自肺炎鏈球菌血清型1、2、3、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15B、18C、19A、19F、22F、23F及33F之醣結合物。在一實施例中，免疫原組合物為22價肺炎鏈球菌結合物組合物。In one embodiment, the present invention relates to an immunogenic composition comprising the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprising saccharide conjugates from pneumococcal serotypes 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F and 33F. In one embodiment, the immunogenic composition is a 22-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含來自肺炎鏈球菌血清型1、2、3、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15C、18C、19A、19F、22F、23F及33F之醣結合物。在一實施例中，免疫原組合物為22價肺炎鏈球菌結合物組合物。In one embodiment, the present invention relates to an immunogenic composition comprising the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprising saccharide conjugates from pneumococcal serotypes 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15C, 18C, 19A, 19F, 22F, 23F and 33F. In one embodiment, the immunogenic composition is a 22-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含來自肺炎鏈球菌血清型1、3、4、5、6A、6B、7F、8、9V、9N、10A、11A、12F、14、15B、18C、19A、19F、22F、23F及33F之醣結合物。在一實施例中，免疫原組合物為22價肺炎鏈球菌結合物組合物。In one embodiment, the present invention relates to an immunogenic composition comprising the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprising saccharide conjugates from pneumococcal serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 9N, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F and 33F. In one embodiment, the immunogenic composition is a 22-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含來自肺炎鏈球菌血清型1、2、3、4、5、6A、6B、7F、8、9V、9N、10A、11A、12F、14、15B、18C、19A、19F、22F、23F及33F之醣結合物。在一實施例中，免疫原組合物為23價肺炎鏈球菌結合物組合物。In one embodiment, the present invention relates to an immunogenic composition comprising the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprising saccharide conjugates from pneumococcal serotypes 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 9N, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F and 33F. In one embodiment, the immunogenic composition is a 23-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含來自肺炎鏈球菌血清型1、2、3、4、5、6A、6B、7F、8、9V、9N、10A、11A、12F、14、15B、17F、18C、19A、19F、22F、23F及33F之醣結合物。在一實施例中，免疫原組合物為24價肺炎鏈球菌結合物組合物。In one embodiment, the present invention relates to an immunogenic composition comprising the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprising saccharide conjugates from pneumococcal serotypes 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 9N, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 22F, 23F and 33F. In one embodiment, the immunogenic composition is a 24-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含來自肺炎鏈球菌血清型1、2、3、4、5、6A、6B、7F、8、9V、9N、10A、11A、12F、14、15B、17F、18C、19A、19F、20、22F、23F及33F之醣結合物。在一實施例中，免疫原組合物為25價肺炎鏈球菌結合物組合物。In one embodiment, the present invention relates to an immunogenic composition comprising the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprising saccharide conjugates from pneumococcal serotypes 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 9N, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F and 33F. In one embodiment, the immunogenic composition is a 25-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含來自肺炎鏈球菌血清型1、3、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15A、15B、18C、19A、19F、22F、23F及33F之醣結合物。在一實施例中，免疫原組合物為22價肺炎鏈球菌結合物組合物。In one embodiment, the present invention relates to an immunogenic composition comprising the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprising saccharide conjugates from pneumococcal serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23F and 33F. In one embodiment, the immunogenic composition is a 22-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含來自肺炎鏈球菌血清型1、3、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15B、18C、19A、19F、22F、23A、23F及33F之醣結合物。在一實施例中，免疫原組合物為22價肺炎鏈球菌結合物組合物。In one embodiment, the present invention relates to an immunogenic composition comprising the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprising saccharide conjugates from pneumococcal serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23A, 23F and 33F. In one embodiment, the immunogenic composition is a 22-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含來自肺炎鏈球菌血清型1、3、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15B、18C、19A、19F、22F、23B、23F及33F之醣結合物。在一實施例中，免疫原組合物為22價肺炎鏈球菌結合物組合物。In one embodiment, the present invention relates to an immunogenic composition comprising the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprising saccharide conjugates from pneumococcal serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23B, 23F and 33F. In one embodiment, the immunogenic composition is a 22-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含來自肺炎鏈球菌血清型1、3、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15B、18C、19A、19F、22F、23F、24F及33F之醣結合物。在一實施例中，免疫原組合物為22價肺炎鏈球菌結合物組合物。In one embodiment, the present invention relates to an immunogenic composition comprising the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprising saccharide conjugates from pneumococcal serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F, 24F and 33F. In one embodiment, the immunogenic composition is a 22-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含來自肺炎鏈球菌血清型1、3、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15B、18C、19A、19F、22F、23F、33F及35B之醣結合物。在一實施例中，免疫原組合物為22價肺炎鏈球菌結合物組合物。In one embodiment, the present invention relates to an immunogenic composition comprising the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprising saccharide conjugates from pneumococcal serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F, 33F and 35B. In one embodiment, the immunogenic composition is a 22-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含來自肺炎鏈球菌血清型1、3、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15A、15B、18C、19A、19F、22F、23A、23F及33F之醣結合物。在一實施例中，免疫原組合物為23價肺炎鏈球菌結合物組合物。In one embodiment, the present invention relates to an immunogenic composition comprising the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprising saccharide conjugates from pneumococcal serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23A, 23F and 33F. In one embodiment, the immunogenic composition is a 23-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含來自肺炎鏈球菌血清型1、3、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15A、15B、18C、19A、19F、22F、23A、23B、23F、24F、33F及35B之醣結合物。在一個實施例中，肺炎鏈球菌醣與CRM 197結合。在一個實施例中，來自血清型1、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15A、15B、18C、19A、19F、22F、23A、23B、23F、24F、33F、35B及38之肺炎鏈球菌醣與CRM 197結合且來自血清型3之肺炎鏈球菌醣與SCP結合。在一實施例中，免疫原組合物為26價肺炎鏈球菌結合物組合物。 In one embodiment, the invention relates to an immunogenic composition comprising the pneumococcal serotype 38 saccharide conjugate of the invention and further comprising saccharide conjugates from pneumococcal serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F and 35B. In one embodiment, the pneumococcal saccharide is conjugated to CRM 197 . In one embodiment, pneumococcal saccharides from serotypes 1, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, 35B and 38 are conjugated to CRM 197 and pneumococcal saccharides from serotype 3 are conjugated to SCP. In one embodiment, the immunogenic composition is a 26-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含至少一種選自由以下組成之群之醣結合物：來自肺炎鏈球菌血清型2、3、7C、9N、10B、15A、16F、17F、20、21、22A、23A、23B、24B、24F、27、29、31、33B、34、35B及35F之醣結合物。In one embodiment, the present invention relates to an immunogenic composition, which comprises the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprises at least one saccharide conjugate selected from the group consisting of: saccharide conjugates from pneumococcal serotypes 2, 3, 7C, 9N, 10B, 15A, 16F, 17F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B and 35F.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含二十一種選自由以下組成之群之醣結合物：來自肺炎鏈球菌血清型2、3、7C、9N、10B、15A、16F、17F、20、21、22A、23A、23B、24B、24F、27、29、31、33B、34、35B及35F之醣結合物。在一實施例中，免疫原組合物為22價肺炎鏈球菌結合物組合物。In one embodiment, the present invention relates to an immunogenic composition, which comprises the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprises twenty-one saccharide conjugates selected from the group consisting of: saccharide conjugates from pneumococcal serotypes 2, 3, 7C, 9N, 10B, 15A, 16F, 17F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B and 35F. In one embodiment, the immunogenic composition is a 22-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含來自血清型2、3、7C、9N、10B、15A、16F、17F、20、21、22A、23A、23B、24B、24F、27、29、31、33B、34、35B及35F之醣結合物。在一實施例中，免疫原組合物為23價肺炎鏈球菌結合物組合物。In one embodiment, the present invention relates to an immunogenic composition comprising the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprising saccharide conjugates from serotypes 2, 3, 7C, 9N, 10B, 15A, 16F, 17F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B and 35F. In one embodiment, the immunogenic composition is a 23-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含至少一種選自由以下組成之群之醣結合物：來自肺炎鏈球菌血清型2、3、7C、9N、10B、15A、16F、17F、19A、19F、20、21、22A、23A、23B、24B、24F、27、29、31、33B、34、35B及35F之醣結合物。In one embodiment, the present invention relates to an immunogenic composition, which comprises the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprises at least one saccharide conjugate selected from the group consisting of: saccharide conjugates from pneumococcal serotypes 2, 3, 7C, 9N, 10B, 15A, 16F, 17F, 19A, 19F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B and 35F.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含二十二種選自由以下組成之群之醣結合物：來自肺炎鏈球菌血清型2、3、7C、9N、10B、15A、16F、17F、19A、19F、20、21、22A、23A、23B、24B、24F、27、29、31、33B、34、35B及35F之醣結合物。在一實施例中，免疫原組合物為23價肺炎鏈球菌結合物組合物。In one embodiment, the present invention relates to an immunogenic composition, which comprises the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprises twenty-two saccharide conjugates selected from the group consisting of saccharide conjugates from pneumococcal serotypes 2, 3, 7C, 9N, 10B, 15A, 16F, 17F, 19A, 19F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B and 35F. In one embodiment, the immunogenic composition is a 23-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含二十三種選自由以下組成之群之醣結合物：來自肺炎鏈球菌血清型2、3、7C、9N、10B、15A、16F、17F、19A、19F、20、21、22A、23A、23B、24B、24F、27、29、31、33B、34、35B及35F之醣結合物。在一實施例中，免疫原組合物為24價肺炎鏈球菌結合物組合物。In one embodiment, the present invention relates to an immunogenic composition comprising the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprising twenty-three saccharide conjugates selected from the group consisting of saccharide conjugates from pneumococcal serotypes 2, 3, 7C, 9N, 10B, 15A, 16F, 17F, 19A, 19F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B and 35F. In one embodiment, the immunogenic composition is a 24-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含來自血清型2、3、7C、9N、10B、15A、16F、17F、20、21、22A、23A、23B、24B、24F、27、29、31、33B、34、35B及35F之醣結合物。在一實施例中，免疫原組合物為23價肺炎鏈球菌結合物組合物。In one embodiment, the present invention relates to an immunogenic composition comprising the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprising saccharide conjugates from serotypes 2, 3, 7C, 9N, 10B, 15A, 16F, 17F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B and 35F. In one embodiment, the immunogenic composition is a 23-valent pneumococcal conjugate composition.

在一實施例中，本發明係關於一種免疫原組合物，其包含本發明之肺炎鏈球菌血清型38醣結合物且其進一步包含來自肺炎鏈球菌血清型2、3、7C、9N、10B、15A、16F、17F、19A、19F、20、21、22A、23A、23B、24B、24F、27、29、31、33B、34、35B及35F之醣結合物。在一實施例中，免疫原組合物為25價肺炎鏈球菌結合物組合物。In one embodiment, the present invention relates to an immunogenic composition comprising the pneumococcal serotype 38 saccharide conjugate of the present invention and further comprising saccharide conjugates from pneumococcal serotypes 2, 3, 7C, 9N, 10B, 15A, 16F, 17F, 19A, 19F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B and 35F. In one embodiment, the immunogenic composition is a 25-valent pneumococcal conjugate composition.

本發明之組合物可包括少量游離載體。當給定載體蛋白以游離及結合形式存在於本發明之組合物中時，非結合形式總體上較佳不超過組合物中之載體蛋白之總量的5%，且更佳以少於2重量%存在。The composition of the present invention may include a small amount of free carrier. When a given carrier protein is present in the composition of the present invention in free and bound forms, the unbound form is generally preferably no more than 5% of the total amount of the carrier protein in the composition, and is more preferably present at less than 2% by weight.

4 佐劑在一些實施例中，本文所揭露之免疫原組合物可進一步包含至少一種、兩種或三種佐劑。在一些實施例中，本文所揭露之免疫原組合物可進一步包含至少一種佐劑。 4 Adjuvants In some embodiments, the immunogenic compositions disclosed herein may further comprise at least one, two or three adjuvants. In some embodiments, the immunogenic compositions disclosed herein may further comprise at least one adjuvant.

在一些實施例中，本文所揭露之免疫原組合物可進一步包含一種佐劑。In some embodiments, the immunogenic compositions disclosed herein may further comprise an adjuvant.

在一些實施例中，本文所揭露之免疫原組合物可進一步包含兩種佐劑。In some embodiments, the immunogenic compositions disclosed herein may further comprise two adjuvants.

術語「佐劑」係指增強針對抗原之免疫反應的化合物或混合物。抗原可主要用作遞送系統，主要用作免疫調節劑，或具有兩者之強力特徵。適合的佐劑包括適用於哺乳動物(包括人類)之彼等佐劑。The term "adjuvant" refers to a compound or mixture that enhances the immune response to an antigen. An antigen may function primarily as a delivery system, primarily as an immunomodulator, or have potent characteristics of both. Suitable adjuvants include those suitable for use in mammals, including humans.

已知可用於人類中之適合的遞送系統類型佐劑之實例包括(但不限於)明礬(例如磷酸鋁、硫酸鋁或氫氧化鋁)、磷酸鈣、脂質體、水包油乳液(諸如MF59)(4.3% w/v角鯊烯、0.5% w/v聚山梨醇酯80 (Tween 80)、0.5% w/v脫水山梨糖醇三油酸酯(Span 85))、油包水乳液(諸如孟塔納(Montanide))及聚(D,L-丙交酯-共-乙交酯) (PLG)微米粒子或奈米粒子。Examples of suitable delivery system types of adjuvants known to be useful in humans include, but are not limited to, alums (e.g., aluminum phosphate, aluminum sulfate, or aluminum hydroxide), calcium phosphate, liposomes, oil-in-water emulsions such as MF59 (4.3% w/v squalene, 0.5% w/v polysorbate 80 (Tween 80), 0.5% w/v sorbitan trioleate (Span 85)), water-in-oil emulsions such as Montanide, and poly(D,L-lactide-co-glycolide) (PLG) microparticles or nanoparticles.

在一實施例中，本文所揭露之免疫原組合物包含作為佐劑之鋁鹽(明礬) (例如磷酸鋁、硫酸鋁或氫氧化鋁)。In one embodiment, the immunogenic compositions disclosed herein comprise an aluminum salt (aluminum) (e.g., aluminum phosphate, aluminum sulfate, or aluminum hydroxide) as an adjuvant.

在一較佳實施例中，本文所揭露之免疫原組合物包含作為佐劑之磷酸鋁或氫氧化鋁。在一甚至較佳實施例中，本文所揭露之免疫原組合物包含作為佐劑之磷酸鋁。In a preferred embodiment, the immunogenic compositions disclosed herein comprise aluminum phosphate or aluminum hydroxide as an adjuvant. In an even more preferred embodiment, the immunogenic compositions disclosed herein comprise aluminum phosphate as an adjuvant.

增強本文所揭露之免疫原組合物之有效性的其他例示性佐劑包括(但不限於)：(1)水包油乳液調配物(含或不含其他特異性免疫刺激劑，諸如胞壁醯基肽(參見下文)或細菌細胞壁組分)，諸如(a) SAF，含有10%角鯊烯、0.4% Tween 80、5% pluronic嵌段聚合物L121及thr-MDP，其微流化成次微米級乳液或渦旋生成較大粒徑的乳液，及(b) RIBI™佐劑系統(RAS)，(Ribi Immunochem, Hamilton, MT)，其含有2%角鯊烯、0.2% Tween 80及一或多種細菌細胞壁組分，諸如單磷醯基脂質A (MPL)、海藻醣二黴菌酸酯(TDM)及細胞壁骨架(CWS)，較佳MPL + CWS (DETOX™)；(2)可使用皂素佐劑，諸如QS21、STIMULON™ (Cambridge Bioscience, Worcester, MA)、ABISCO® (Isconova, Sweden)或ISCOMATRIX® (Commonwealth Serum Laboratories, Australia)，或由其產生之粒子，諸如免疫刺激複合物(ISCOM)，該ISCOM可不含額外清潔劑(例如WO 00/07621)；(3)弗氏完全佐劑(Complete Freund's Adjuvant) (CFA)及弗氏不完全佐劑(Incomplete Freund's Adjuvant) (IFA)；(4)細胞介素，諸如介白素(例如IL-1、IL-2、IL-4、IL-5、IL-6、IL-7、IL-12 (例如WO 99/44636))、干擾素(例如γ干擾素)、巨噬細胞群落刺激因子(M-CSF)、腫瘤壞死因子(TNF)等；(5)單磷醯基脂質A (MPL)或3-O-去醯化MPL (3dMPL) (參見例如GB-2220221、EP0689454)，視情況在與肺炎鏈球菌醣一起使用時實質上不存在明礬(參見例如WO 00/56358)；(6) 3dMPL與例如QS21及/或水包油乳液之組合(參見例如EP0835318、EP0735898、EP0761231)；(7)聚氧化乙烯醚或聚氧化乙烯酯(參見例如WO 99/52549)；(8)與辛苯聚醇組合之聚氧化乙烯脫水山梨糖醇酯界面活性劑(例如WO 01/21207)或與至少一種額外非離子界面活性劑(諸如辛苯聚醇)組合之聚氧化乙烯烷基醚或酯界面活性劑(例如WO 01/21152)；(9)皂素及免疫刺激性寡核苷酸(例如CpG寡核苷酸) (例如WO 00/62800)；(10)免疫刺激劑及金屬鹽之粒子(參見例如WO 00/23105)；(11)皂素及水包油乳液(例如WO 99/11241)；(12)皂素(例如QS21) + 3dMPL + IM2 (視情況+固醇) (例如WO 98/57659)；(13)充當免疫刺激劑以增強組合物之功效的其他物質。胞壁醯基肽包括N-乙醯基-胞壁醯基-L-羥丁胺醯基-D-異麩醯胺酸(thr-MDP)、N-25乙醯基-去甲胞壁醯基-L-丙胺醯基-D-異麩醯胺酸(nor-MDP)、N-乙醯基胞壁醯基-L-丙胺醯基-D-異麩醯胺基-L-丙胺酸-2-(1'-2'-二軟脂醯基-sn-丙三氧基-3-羥基磷醯基氧基)-乙胺MTP-PE)等。Other exemplary adjuvants that enhance the effectiveness of the immunogenic compositions disclosed herein include, but are not limited to: (1) oil-in-water emulsion formulations (with or without other specific immunostimulants, such as cell wall acyl peptides (see below) or bacterial cell wall components), such as (a) SAF, containing 10% squalene, 0.4% Tween 80, 5% pluronic block polymer L121 and thr-MDP, which is microfluidized into a submicron emulsion or vortexed to form an emulsion with larger particle size, and (b) RIBI™ Adjuvant System (RAS), (Ribi Immunochem, Hamilton, MT), which contains 2% squalene, 0.2% Tween 80 and one or more bacterial cell wall components, such as monophosphoryl lipid A (MPL), trehalose dimelatin (TDM) and cell wall skeleton (CWS), preferably MPL + CWS (DETOX™); (2) saponin adjuvants such as QS21, STIMULON™ (Cambridge Bioscience, Worcester, MA), ABISCO® (Isconova, Sweden) or ISCOMATRIX® (Commonwealth Serum Laboratories, Australia), or particles derived therefrom such as immunostimulatory complexes (ISCOMs), which may not contain additional detergents (e.g., WO 00/07621); (3) Complete Freund's Adjuvant (CFA) and Incomplete Freund's Adjuvant (CFA) (IFA); (4) interleukins, such as interleukins (e.g., IL-1, IL-2, IL-4, IL-5, IL-6, IL-7, IL-12 (e.g., WO 99/44636)), interferons (e.g., gamma interferon), macrophage colony stimulating factor (M-CSF), tumor necrosis factor (TNF), etc.; (5) monophosphoryl lipid A (MPL) or 3-O-deacylated MPL (3dMPL) (see, e.g., GB-2220221, EP0689454), which are substantially free of alum when used together with pneumococcal saccharide (see, e.g., WO 00/56358); (6) 3dMPL in combination with, for example, QS21 and/or an oil-in-water emulsion (see, for example, EP0835318, EP0735898, EP0761231); (7) polyoxyethylene ethers or polyoxyethylene esters (see, for example, WO 99/52549); (8) polyoxyethylene sorbitan ester surfactants in combination with octoxynol (e.g., WO 01/21207) or polyoxyethylene alkyl ether or ester surfactants in combination with at least one additional non-ionic surfactant (such as octoxynol) (e.g., WO 01/21152); (9) saponin and immunostimulatory oligonucleotides (e.g., CpG oligonucleotides) (e.g., WO 00/62800); (10) particles of immunostimulatory agents and metal salts (see, for example, WO 00/23105); (11) saponin and oil-in-water emulsions (e.g. WO 99/11241); (12) saponin (e.g. QS21) + 3dMPL + IM2 (optionally + steroids) (e.g. WO 98/57659); (13) other substances that act as immunostimulants to enhance the efficacy of the composition. The muramyl peptides include N-acetyl-muramyl-L-hydroxybutylamino-D-isoglutamine (thr-MDP), N-25-acetyl-normuramyl-L-propylamino-D-isoglutamine (nor-MDP), N-acetylmuramyl-L-propylamino-D-isoglutamine-L-alanine-2-(1'-2'-dimaltoyl-sn-glycerotrioxy-3-hydroxyphosphoyloxy)-ethylamine (MTP-PE), and the like.

在本發明之一實施例中，如本文所揭露之免疫原組合物包含作為佐劑之CpG寡核苷酸。如本文中所使用之CpG寡核苷酸係指免疫刺激性CpG寡去氧核苷酸(CpG ODN)，且因此除非另外指示，否則此等術語可互換使用。免疫刺激性CpG寡去氧核苷酸含有一或多個免疫刺激性CPG模體，其為未甲基化胞嘧啶-鳥嘌呤二核苷酸，視情況在某些較佳的鹼基背景內。CpG免疫刺激性模體之甲基化狀態通常係指二核苷酸中之胞嘧啶殘基。含有至少一個未甲基化CpG二核苷酸之免疫刺激性寡核苷酸為含有藉由磷酸酯鍵與3'鳥嘌呤連接之5'未甲基化胞嘧啶且透過與鐸樣受體9 (TLR-9)結合而活化免疫系統的寡核苷酸。在另一實施例中，免疫刺激性寡核苷酸可含有一或多個甲基化CpG二核苷酸，其將透過TLR9活化免疫系統但不如CpG模體之未甲基化一樣強烈。CpG免疫刺激性寡核苷酸可包含一或多個回文序列，其亦可涵蓋CpG二核苷酸。CpG寡核苷酸已在多個發佈專利、公開專利申請案及其他公開案中描述，包括美國專利第6,194,388號、第6,207,646號、第6,214,806號、第6,218,371號、第6,239,116號；及第6,339,068號。In one embodiment of the present invention, the immunogenic composition as disclosed herein comprises a CpG oligonucleotide as an adjuvant. As used herein, a CpG oligonucleotide refers to an immunostimulatory CpG oligodeoxynucleotide (CpG ODN), and therefore unless otherwise indicated, these terms are used interchangeably. An immunostimulatory CpG oligodeoxynucleotide contains one or more immunostimulatory CpG motifs, which are unmethylated cytosine-guanine dinucleotides, optionally in certain preferred alkaline contexts. The methylation state of a CpG immunostimulatory motif generally refers to the cytosine residue in the dinucleotide. An immunostimulatory oligonucleotide containing at least one unmethylated CpG dinucleotide is an oligonucleotide containing a 5' unmethylated cytosine linked to a 3' guanine by a phosphate bond and activating the immune system by binding to toll-like receptor 9 (TLR-9). In another embodiment, the immunostimulatory oligonucleotide may contain one or more methylated CpG dinucleotides, which will activate the immune system through TLR9 but not as strongly as the unmethylated CpG motif. The CpG immunostimulatory oligonucleotide may comprise one or more palindromic sequences, which may also encompass CpG dinucleotides. CpG oligonucleotides have been described in a number of issued patents, published patent applications, and other publications, including U.S. Patent Nos. 6,194,388, 6,207,646, 6,214,806, 6,218,371, 6,239,116; and 6,339,068.

在本發明之一實施例中，如本文所揭露之免疫原組合物包含WO 2010/125480之第3頁第22行至第12頁第36行所描述之CpG寡核苷酸中之任一者。In one embodiment of the present invention, the immunogenic composition as disclosed herein comprises any one of the CpG oligonucleotides described on page 3, line 22 to page 12, line 36 of WO 2010/125480.

已鑑別出不同類別之CpG免疫刺激性寡核苷酸。此等被稱為A、B、C及P類，且更詳細地描述於WO 2010/125480之第3頁第22行至第12頁第36行。本發明之方法涵蓋此等不同類別之CpG免疫刺激性寡核苷酸之用途。Different classes of CpG immunostimulatory oligonucleotides have been identified. These are referred to as classes A, B, C and P, and are described in more detail in WO 2010/125480, page 3, line 22 to page 12, line 36. The methods of the present invention encompass the use of these different classes of CpG immunostimulatory oligonucleotides.

5 調配物本發明之免疫原組合物可調配呈液體形式(亦即溶液或懸浮液)或呈凍乾形式。在一實施例中，本發明之免疫原組合物調配成液體形式。在一實施例中，本發明之免疫原組合物調配成凍乾形式。液體調配物可有利地自其封裝形式直接投與，且因此對於注射而言為理想的，其無需如本發明之凍乾組合物那般在水性介質中復原。 5 Formulations The immunogenic compositions of the present invention may be formulated in liquid form (i.e., solutions or suspensions) or in lyophilized form. In one embodiment, the immunogenic compositions of the present invention are formulated in liquid form. In one embodiment, the immunogenic compositions of the present invention are formulated in lyophilized form. Liquid formulations can advantageously be administered directly from their encapsulated form and are therefore ideal for injection, without the need for reconstitution in an aqueous medium as is the case with the lyophilized compositions of the present invention.

本揭露之免疫原組合物之調配可使用此項技術中公認的方法來進行。舉例而言，可將個別多醣及/或結合物與生理學上可接受之媒劑一起調配以製備組合物。此類媒劑之實例包括(但不限於)水、緩衝鹽水、多元醇(例如，甘油、丙二醇、液體聚乙二醇)及右旋糖溶液。The immunogenic compositions disclosed herein can be formulated using methods generally recognized in the art. For example, individual polysaccharides and/or conjugates can be formulated with physiologically acceptable vehicles to prepare the compositions. Examples of such vehicles include, but are not limited to, water, buffered saline, polyols (e.g., glycerol, propylene glycol, liquid polyethylene glycol), and dextrose solutions.

本揭露提供一種免疫原組合物，其包含本文所揭露之醣結合物與醫藥學上可接受之賦形劑、載劑或稀釋劑之組合中之任一者。The present disclosure provides an immunogenic composition comprising any one of the saccharide conjugates disclosed herein in combination with a pharmaceutically acceptable excipient, carrier or diluent.

在一實施例中，本揭露之免疫原組合物呈液體形式，較佳地呈水性液體形式。In one embodiment, the immunogenic composition of the present disclosure is in liquid form, preferably in aqueous liquid form.

本揭露之免疫原組合物可包含以下中之一或多者：緩衝液、鹽、二價陽離子、非離子清潔劑、低溫保護劑(諸如糖)及抗氧化劑(諸如游離基清除劑或螯合劑)或其任何多個組合。The immunogenic composition disclosed herein may include one or more of the following: a buffer, a salt, a divalent cation, a non-ionic detergent, a low-temperature protectant (such as sugar) and an antioxidant (such as a free radical scavenger or a chelating agent) or any combination thereof.

在一實施例中，本揭露之免疫原組合物包含緩衝液。在一實施例中，該緩衝液之pKa為約3.5至約7.5。在一些實施例中，緩衝劑為磷酸鹽、丁二酸鹽、組胺酸或檸檬酸鹽。在一些實施例中，緩衝液為丁二酸鹽。在一些實施例中，緩衝液為組胺酸。在某些實施例中，緩衝液為在1 mM至10 mM之最終濃度下之丁二酸鹽。在一個特定實施例中，丁二酸鹽緩衝液之最終濃度為約5 mM。In one embodiment, the immunogenic composition of the present disclosure comprises a buffer. In one embodiment, the pKa of the buffer is about 3.5 to about 7.5. In some embodiments, the buffer is phosphate, succinate, histidine, or citrate. In some embodiments, the buffer is succinate. In some embodiments, the buffer is histidine. In certain embodiments, the buffer is succinate at a final concentration of 1 mM to 10 mM. In a specific embodiment, the final concentration of the succinate buffer is about 5 mM.

在一實施例中，本揭露之免疫原組合物包含鹽。在一些實施例中，鹽係選自由以下組成之群：氯化鎂、氯化鉀、氯化鈉及其組合。在一個特定實施例中，鹽為氯化鈉。在一個特定實施例中，本發明之免疫原組合物包含150 mM之氯化鈉。In one embodiment, the immunogenic composition of the present disclosure comprises a salt. In some embodiments, the salt is selected from the group consisting of magnesium chloride, potassium chloride, sodium chloride, and combinations thereof. In a specific embodiment, the salt is sodium chloride. In a specific embodiment, the immunogenic composition of the present invention comprises 150 mM sodium chloride.

在一實施例中，本揭露之免疫原組合物包含界面活性劑。在一實施例中，界面活性劑係選自由以下組成之群：聚山梨醇酯20 (TWEEN TM20)、聚山梨醇酯40 (TWEEN TM40)、聚山梨醇酯60 (TWEEN TM60)、聚山梨醇酯65 (TWEEN TM65)、聚山梨醇酯80 (TWEEN TM80)、聚山梨醇酯85 (TWEEN TM85)、TRITON TMN-101、TRITON TMX-100、辛苯聚醇40、壬苯醇醚-9、三乙醇胺、三乙醇胺多肽油酸酯、聚氧化乙烯-660羥基硬脂酸酯(PEG-15、Solutol H 15)、聚氧化乙烯-35-蓖麻油酸酯(CREMOPHOR® EL)、大豆卵磷脂及泊洛沙姆(poloxamer)。 In one embodiment, the immunogenic composition of the present disclosure comprises a surfactant. In one embodiment, the surfactant is selected from the group consisting of polysorbate 20 (TWEEN ™ 20), polysorbate 40 (TWEEN ™ 40), polysorbate 60 (TWEEN ™ 60), polysorbate 65 (TWEEN ™ 65), polysorbate 80 (TWEEN ™ 80), polysorbate 85 (TWEEN ™ 85), TRITON ™ N-101, TRITON ™ X-100, octoxynol 40, nonoxynol-9, triethanolamine, triethanolamine polypeptide oleate, polyoxyethylene-660 hydroxystearate (PEG-15, Solutol H 15), polyoxyethylene-35-ricinoleate (CREMOPHOR® EL), soy lecithin and poloxamer.

在一個較佳實施例中，界面活性劑為聚山梨醇酯80。在一些該實施例中，調配物中聚山梨醇酯80之最終濃度為至少0.0001%至10% (重量/重量) (w/w)聚山梨醇酯80。在一些該等實施例中，調配物中聚山梨醇酯80之最終濃度為至少0.001%至1%重量/重量(w/w)之聚山梨醇酯80。在一些該等實施例中，調配物中聚山梨醇酯80之最終濃度為至少0.01%至1%重量/重量(w/w)之聚山梨醇酯80。在其他實施例中，調配物中聚山梨醇酯80之最終濃度為0.01%、0.02%、0.03%、0.04%、0.05%、0.06%、0.07%、0.08%、0.09%或0.1% (w/w)聚山梨醇酯80。在另一實施例中，調配物中聚山梨醇酯80之最終濃度為0.02% (w/w)聚山梨醇酯80。在另一實施例中，調配物中聚山梨醇酯80之最終濃度為0.01% (w/w)聚山梨醇酯80。在另一實施例中，調配物中聚山梨醇酯80之最終濃度為0.03% (w/w)聚山梨醇酯80。在另一實施例中，調配物中聚山梨醇酯80之最終濃度為0.04% (w/w)聚山梨醇酯80。在另一實施例中，調配物中聚山梨醇酯80之最終濃度為0.05% (w/w)聚山梨醇酯80。在另一實施例中，調配物中聚山梨醇酯80之最終濃度為1% (w/w)聚山梨醇酯80。In a preferred embodiment, the surfactant is polysorbate 80. In some of these embodiments, the final concentration of polysorbate 80 in the formulation is at least 0.0001% to 10% (weight/weight) (w/w) polysorbate 80. In some of these embodiments, the final concentration of polysorbate 80 in the formulation is at least 0.001% to 1% weight/weight (w/w) polysorbate 80. In some of these embodiments, the final concentration of polysorbate 80 in the formulation is at least 0.01% to 1% weight/weight (w/w) polysorbate 80. In other embodiments, the final concentration of polysorbate 80 in the formulation is 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09% or 0.1% (w/w) polysorbate 80. In another embodiment, the final concentration of polysorbate 80 in the formulation is 0.02% (w/w) polysorbate 80. In another embodiment, the final concentration of polysorbate 80 in the formulation is 0.01% (w/w) polysorbate 80. In another embodiment, the final concentration of polysorbate 80 in the formulation is 0.03% (w/w) polysorbate 80. In another embodiment, the final concentration of polysorbate 80 in the formulation is 0.04% (w/w) polysorbate 80. In another embodiment, the final concentration of polysorbate 80 in the formulation is 0.05% (w/w) polysorbate 80. In another embodiment, the final concentration of polysorbate 80 in the formulation is 1% (w/w) polysorbate 80.

在一個特定實施例中，界面活性劑為聚山梨醇酯20。在一些該實施例中，調配物中聚山梨醇酯20之最終濃度為至少0.0001%至10%重量/重量(w/w)聚山梨醇酯20。在一些該等實施例中，調配物中聚山梨醇酯20之最終濃度為至少0.001%至1%重量/重量(w/w)聚山梨醇酯20。在一些該等實施例中，調配物中之聚山梨醇酯20之最終濃度為至少0.01%至1%重量/重量(w/w)聚山梨醇酯20。在其他實施例中，調配物中聚山梨醇酯20之最終濃度為0.01%、0.02%、0.03%、0.04%、0.05%、0.06%、0.07%、0.08%、0.09%或0.1% (w/w)聚山梨醇酯20。在另一實施例中，調配物中聚山梨醇酯20之最終濃度為0.02% (w/w)聚山梨醇酯20。在另一實施例中，調配物中聚山梨醇酯20之最終濃度為0.01% (w/w)聚山梨醇酯20。在另一實施例中，調配物中聚山梨醇酯20之最終濃度為0.03% (w/w)聚山梨醇酯20。在另一實施例中，調配物中聚山梨醇酯20之最終濃度為0.04% (w/w)聚山梨醇酯80。在另一實施例中，調配物中聚山梨醇酯20之最終濃度為0.05% (w/w)聚山梨醇酯20。在另一實施例中，調配物中聚山梨醇酯20之最終濃度為1% (w/w)聚山梨醇酯20。In a particular embodiment, the surfactant is polysorbate 20. In some of these embodiments, the final concentration of polysorbate 20 in the formulation is at least 0.0001% to 10% weight/weight (w/w) polysorbate 20. In some of these embodiments, the final concentration of polysorbate 20 in the formulation is at least 0.001% to 1% weight/weight (w/w) polysorbate 20. In some of these embodiments, the final concentration of polysorbate 20 in the formulation is at least 0.01% to 1% weight/weight (w/w) polysorbate 20. In other embodiments, the final concentration of polysorbate 20 in the formulation is 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, or 0.1% (w/w) polysorbate 20. In another embodiment, the final concentration of polysorbate 20 in the formulation is 0.02% (w/w) polysorbate 20. In another embodiment, the final concentration of polysorbate 20 in the formulation is 0.01% (w/w) polysorbate 20. In another embodiment, the final concentration of polysorbate 20 in the formulation is 0.03% (w/w) polysorbate 20. In another embodiment, the final concentration of polysorbate 20 in the formulation is 0.04% (w/w) polysorbate 80. In another embodiment, the final concentration of polysorbate 20 in the formulation is 0.05% (w/w) polysorbate 20. In another embodiment, the final concentration of polysorbate 20 in the formulation is 1% (w/w) polysorbate 20.

在一個特定實施例中，界面活性劑為聚山梨醇酯40。在某一該實施例中，調配物中聚山梨醇酯40之最終濃度為至少0.0001%至10%重量/重量(w/w)之聚山梨醇酯40。在一些該等實施例中，調配物中聚山梨醇酯40之最終濃度為至少0.001%至1%重量/重量(w/w)之聚山梨醇酯40。在一些該等實施例中，調配物中聚山梨醇酯40之最終濃度為至少0.01%至1%重量/重量(w/w)之聚山梨醇酯40。在其他實施例中，調配物中聚山梨醇酯40之最終濃度為0.01%、0.02%、0.03%、0.04%、0.05%、0.06%、0.07%、0.08%、0.09%或0.1% (w/w)聚山梨醇酯40。在另一實施例中，調配物中聚山梨醇酯40之最終濃度為1% (w/w)聚山梨醇酯40。In a particular embodiment, the surfactant is polysorbate 40. In a certain such embodiment, the final concentration of polysorbate 40 in the formulation is at least 0.0001% to 10% weight/weight (w/w) polysorbate 40. In some such embodiments, the final concentration of polysorbate 40 in the formulation is at least 0.001% to 1% weight/weight (w/w) polysorbate 40. In some such embodiments, the final concentration of polysorbate 40 in the formulation is at least 0.01% to 1% weight/weight (w/w) polysorbate 40. In other embodiments, the final concentration of polysorbate 40 in the formulation is 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09% or 0.1% (w/w) polysorbate 40. In another embodiment, the final concentration of polysorbate 40 in the formulation is 1% (w/w) polysorbate 40.

在一個特定實施例中，界面活性劑為聚山梨醇酯60。在某一該實施例中，調配物中聚山梨醇酯60之最終濃度為至少0.0001%至10%重量/重量(w/w)之聚山梨醇酯60。在一些該等實施例中，調配物中聚山梨醇酯60之最終濃度為至少0.001%至1%重量/重量(w/w)之聚山梨醇酯60。在一些該等實施例中，調配物中聚山梨醇酯60之最終濃度為至少0.01%至1%重量/重量(w/w)之聚山梨醇酯60。在其他實施例中，調配物中聚山梨醇酯60之最終濃度為0.01%、0.02%、0.03%、0.04%、0.05%、0.06%、0.07%、0.08%、0.09%或0.1% (w/w)聚山梨醇酯60。在另一實施例中，調配物中聚山梨醇酯60之最終濃度為1% (w/w)聚山梨醇酯60。In a particular embodiment, the surfactant is polysorbate 60. In a certain such embodiment, the final concentration of polysorbate 60 in the formulation is at least 0.0001% to 10% weight/weight (w/w) polysorbate 60. In some such embodiments, the final concentration of polysorbate 60 in the formulation is at least 0.001% to 1% weight/weight (w/w) polysorbate 60. In some such embodiments, the final concentration of polysorbate 60 in the formulation is at least 0.01% to 1% weight/weight (w/w) polysorbate 60. In other embodiments, the final concentration of polysorbate 60 in the formulation is 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09% or 0.1% (w/w) polysorbate 60. In another embodiment, the final concentration of polysorbate 60 in the formulation is 1% (w/w) polysorbate 60.

在一個特定實施例中，界面活性劑為聚山梨醇酯65。在某一該實施例中，調配物中聚山梨醇酯65之最終濃度為至少0.0001%至10%重量/重量(w/w)之聚山梨醇酯65。在一些該等實施例中，調配物中聚山梨醇酯65之最終濃度為至少0.001%至1%重量/重量(w/w)之聚山梨醇酯65。在一些該等實施例中，調配物中聚山梨醇酯65之最終濃度為至少0.01%至1%重量/重量(w/w)之聚山梨醇酯65。在其他實施例中，調配物中聚山梨醇酯65之最終濃度為0.01%、0.02%、0.03%、0.04%、0.05%、0.06%、0.07%、0.08%、0.09%或0.1% (w/w)聚山梨醇酯65。在另一實施例中，調配物中聚山梨醇酯65之最終濃度為1% (w/w)聚山梨醇酯65。In a particular embodiment, the surfactant is polysorbate 65. In a certain such embodiment, the final concentration of polysorbate 65 in the formulation is at least 0.0001% to 10% weight/weight (w/w) polysorbate 65. In some such embodiments, the final concentration of polysorbate 65 in the formulation is at least 0.001% to 1% weight/weight (w/w) polysorbate 65. In some such embodiments, the final concentration of polysorbate 65 in the formulation is at least 0.01% to 1% weight/weight (w/w) polysorbate 65. In other embodiments, the final concentration of polysorbate 65 in the formulation is 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09% or 0.1% (w/w) polysorbate 65. In another embodiment, the final concentration of polysorbate 65 in the formulation is 1% (w/w) polysorbate 65.

在一個特定實施例中，界面活性劑為聚山梨醇酯85。在某一該實施例中，調配物中聚山梨醇酯85之最終濃度為至少0.0001%至10%重量/重量(w/w)之聚山梨醇酯85。在一些該等實施例中，調配物中聚山梨醇酯85之最終濃度為至少0.001%至1%重量/重量(w/w)之聚山梨醇酯85。在一些該等實施例中，調配物中聚山梨醇酯85之最終濃度為至少0.01%至1%重量/重量(w/w)之聚山梨醇酯85。在其他實施例中，調配物中聚山梨醇酯85之最終濃度為0.01%、0.02%、0.03%、0.04%、0.05%、0.06%、0.07%、0.08%、0.09%或0.1% (w/w)聚山梨醇酯85。在另一實施例中，調配物中聚山梨醇酯85之最終濃度為1% (w/w)聚山梨醇酯85。In a particular embodiment, the surfactant is polysorbate 85. In a certain such embodiment, the final concentration of polysorbate 85 in the formulation is at least 0.0001% to 10% weight/weight (w/w) of polysorbate 85. In some such embodiments, the final concentration of polysorbate 85 in the formulation is at least 0.001% to 1% weight/weight (w/w) of polysorbate 85. In some such embodiments, the final concentration of polysorbate 85 in the formulation is at least 0.01% to 1% weight/weight (w/w) of polysorbate 85. In other embodiments, the final concentration of polysorbate 85 in the formulation is 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09% or 0.1% (w/w) polysorbate 85. In another embodiment, the final concentration of polysorbate 85 in the formulation is 1% (w/w) polysorbate 85.

在某些實施例中，本揭露之免疫原組合物具有5.5至7.5之pH,更佳5.6至7.0之pH,甚至更佳5.8至6.0之pH。In certain embodiments, the immunogenic compositions of the present disclosure have a pH of 5.5 to 7.5, more preferably a pH of 5.6 to 7.0, and even more preferably a pH of 5.8 to 6.0.

在一個實施例中，本揭露提供一種填充有本文所揭露之免疫原組合物中之任一者的容器。在一個實施例中，容器係選自由以下組成之群：小瓶、注射器、燒瓶、醱酵器、生物反應器、袋子、廣口瓶、安瓿、藥筒及拋棄式筆。在某些實施例中，容器為矽化的。In one embodiment, the disclosure provides a container filled with any of the immunogenic compositions disclosed herein. In one embodiment, the container is selected from the group consisting of a vial, a syringe, a flask, a fermenter, a bioreactor, a bag, a jar, an ampoule, a cartridge, and a disposable pen. In certain embodiments, the container is siliconized.

在一實施例中，本揭露之容器係由玻璃、金屬(例如鋼、不鏽鋼、鋁等)及/或聚合物(例如熱塑性塑膠、彈性體、熱塑性彈性體)製成。在一實施例中，本揭露之容器係由玻璃製成。In one embodiment, the container of the present disclosure is made of glass, metal (e.g., steel, stainless steel, aluminum, etc.) and/or polymer (e.g., thermoplastic plastic, elastomer, thermoplastic elastomer). In one embodiment, the container of the present disclosure is made of glass.

在一個實施例中，本揭露提供一種填充有本文所揭露之免疫原組合物中之任一者的注射器。在某些實施例中，注射器為矽化的及/或係由玻璃製成。In one embodiment, the disclosure provides a syringe filled with any of the immunogenic compositions disclosed herein. In certain embodiments, the syringe is siliconized and/or made of glass.

用於注射之本發明之免疫原組合物的典型劑量具有0.1 mL至2 mL之體積。在一實施例中，用於注射之本發明之免疫原組合物具有0.2 mL至1 mL之體積，甚至更佳約0.5 mL之體積。A typical dose of the immunogenic composition of the present invention for injection has a volume of 0.1 mL to 2 mL. In one embodiment, the immunogenic composition of the present invention for injection has a volume of 0.2 mL to 1 mL, and even more preferably a volume of about 0.5 mL.

6 本發明之醣結合物及免疫原組合物的用途本文所揭露之醣結合物可用作抗原。舉例而言，其可為疫苗之一部分。 6 Uses of the Carbohydrate Conjugates and Immunogenic Compositions of the Invention The carbohydrate conjugates disclosed herein can be used as antigens. For example, they can be part of a vaccine.

因此，在一實施例中，本發明之免疫原組合物適用作藥劑。Therefore, in one embodiment, the immunogenic compositions of the present invention are suitable for use as a medicament.

在一實施例中，本發明之免疫原組合物適用作疫苗。In one embodiment, the immunogenic compositions of the present invention are suitable for use as vaccines.

因此，在一實施例中，本文所描述之免疫原組合物適用於在個體中產生免疫反應。在一個態樣中，個體為哺乳動物，諸如人類、非人類靈長類動物、貓、綿羊、豬、馬、牛科動物或狗。較佳地，個體為人類。Thus, in one embodiment, the immunogenic compositions described herein are suitable for generating an immune response in an individual. In one aspect, the individual is a mammal, such as a human, a non-human primate, a cat, a sheep, a pig, a horse, a bovine, or a dog. Preferably, the individual is a human.

本文所描述之免疫原組合物可用於預防、治療或改善個體之細菌性感染、疾病或病況之治療性或預防性方法中。特定言之，本文所描述之免疫原組合物可用於預防、治療或改善個體之肺炎鏈球菌感染、疾病或病況。較佳地，本文所描述之免疫原組合物可藉由組合物中所含之血清型用於預防、治療或改善個體之肺炎鏈球菌感染、疾病或病況。The immunogenic compositions described herein can be used in therapeutic or prophylactic methods for preventing, treating or ameliorating bacterial infections, diseases or conditions in individuals. Specifically, the immunogenic compositions described herein can be used to prevent, treat or ameliorate pneumococcal infections, diseases or conditions in individuals. Preferably, the immunogenic compositions described herein can be used to prevent, treat or ameliorate pneumococcal infections, diseases or conditions in individuals by virtue of the serotypes contained in the compositions.

因此，在一個態樣中，本揭露提供一種預防、治療或改善個體之與肺炎鏈球菌血清型38相關之感染、疾病或病況的方法，其包含向該個體投與免疫有效量之本揭露之免疫原組合物。Therefore, in one aspect, the present disclosure provides a method for preventing, treating or ameliorating an infection, disease or condition associated with S. pneumoniae serotype 38 in a subject, comprising administering to the subject an immunogenic composition of the present disclosure in an immunologically effective amount.

在一些此類實施例中，該感染、疾病或病況係選自由以下組成之群：肺炎、鼻竇炎、中耳炎、急性中耳炎、腦膜炎、菌血症、敗血症、胸膜積膿、結膜炎、骨髓炎、感染性關節炎、心內膜炎、腹膜炎、心包膜炎、乳突炎、蜂窩組織炎、軟組織感染及大腦膿腫。In some such embodiments, the infection, disease or condition is selected from the group consisting of pneumonia, sinusitis, otitis media, acute otitis media, meningitis, bacteremia, sepsis, pleural effusion, conjunctivitis, osteomyelitis, infectious arthritis, endocarditis, peritonitis, pericarditis, mastoiditis, cellulitis, soft tissue infection and cerebral abscess.

在一實施例中，本揭露提供一種誘導個體中之針對肺炎鏈球菌血清型38之免疫反應的方法，其包含向該個體投與免疫有效量之本發明之免疫原組合物。在一個態樣中，個體為哺乳動物，諸如人類、貓、綿羊、豬、馬、牛科動物或狗。較佳地，個體為人類。In one embodiment, the present disclosure provides a method of inducing an immune response against S. pneumoniae serotype 38 in a subject, comprising administering to the subject an immunogenic composition of the present invention in an immunologically effective amount. In one aspect, the subject is a mammal, such as a human, cat, sheep, pig, horse, bovine, or dog. Preferably, the subject is a human.

在一實施例中，本文所揭露之免疫原組合物適用作疫苗。在此類實施例中，本文所描述之免疫原組合物可用於預防個體中之肺炎鏈球菌血清型38感染。因此，在一個態樣中，本發明提供一種預防個體中受肺炎鏈球菌38感染之方法，其包含向該個體投與免疫有效量之本揭露之免疫原組合物。在一些此類實施例中，該感染係選自由以下組成之群：肺炎、鼻竇炎、中耳炎、急性中耳炎、腦膜炎、菌血症、敗血症、胸膜積膿、結膜炎、骨髓炎、感染性關節炎、心內膜炎、腹膜炎、心包膜炎、乳突炎、蜂窩組織炎、軟組織感染及大腦膿腫。在一個態樣中，個體為哺乳動物，諸如人類、貓、綿羊、豬、馬、牛科動物或狗。較佳地，個體為人類。In one embodiment, the immunogenic compositions disclosed herein are suitable for use as vaccines. In such embodiments, the immunogenic compositions described herein can be used to prevent infection with S. pneumoniae serotype 38 in an individual. Thus, in one aspect, the present invention provides a method for preventing infection with S. pneumoniae serotype 38 in an individual, comprising administering to the individual an immunogenic composition of the present disclosure in an immunologically effective amount. In some such embodiments, the infection is selected from the group consisting of pneumonia, sinusitis, otitis media, acute otitis media, meningitis, bacteremia, septicemia, pleural effusion, conjunctivitis, osteomyelitis, infectious arthritis, endocarditis, peritonitis, pericarditis, mastoiditis, cellulitis, soft tissue infection, and cerebral abscess. In one aspect, the subject is a mammal, such as a human, a cat, a sheep, a pig, a horse, a bovine, or a dog. Preferably, the subject is a human.

本揭露之免疫原組合物可用於保護或治療易受肺炎鏈球菌血清型38感染影響之人類，其係藉助於經由全身或黏膜途徑來投與該免疫原組合物。在一實施例中，本發明之免疫原組合物係藉由肌肉內、腹膜內、皮內或皮下途徑投與。在一實施例中，本發明之免疫原組合物係藉由肌肉內、腹膜內、皮內或皮下注射投與。在一實施例中，本發明之免疫原組合物係藉由肌肉內或皮下注射投與。在一實施例中，本發明之免疫原組合物係藉由肌肉內注射投與。在一實施例中，本發明之免疫原組合物係藉由皮下注射投與。The immunogenic compositions disclosed herein can be used to protect or treat humans susceptible to infection with S. pneumoniae serotype 38 by administering the immunogenic compositions via a systemic or mucosal route. In one embodiment, the immunogenic compositions of the present invention are administered by intramuscular, intraperitoneal, intradermal, or subcutaneous routes. In one embodiment, the immunogenic compositions of the present invention are administered by intramuscular, intraperitoneal, intradermal, or subcutaneous injection. In one embodiment, the immunogenic compositions of the present invention are administered by intramuscular or subcutaneous injection. In one embodiment, the immunogenic compositions of the present invention are administered by intramuscular injection. In one embodiment, the immunogenic compositions of the present invention are administered by subcutaneous injection.

7 待用本發明之免疫原組合物治療的個體如本文所揭露，本文所描述之免疫原組合物可用於預防、治療或改善個體之細菌性感染、疾病或病況之各種治療性或預防性方法中。 7 Subjects to be Treated with the Immunogenic Compositions of the Invention As disclosed herein, the immunogenic compositions described herein can be used in various therapeutic or prophylactic methods for preventing, treating or ameliorating bacterial infections, diseases or conditions in subjects.

在一較佳實施例中，該個體為人類。在一個最佳實施例中，該個體為新生兒(亦即低於三月齡)、嬰兒(亦即年齡在3個月至一歲)或幼兒(亦即一歲至四歲)。在一實施例中，本文所揭露之免疫原組合物適用作疫苗。In a preferred embodiment, the subject is a human. In a most preferred embodiment, the subject is a newborn (i.e., less than three months old), an infant (i.e., aged between three months and one year old), or a toddler (i.e., one to four years old). In one embodiment, the immunogenic compositions disclosed herein are suitable for use as vaccines.

在此類實施例中，待進行疫苗接種之個體可小於1歲。舉例而言，待進行疫苗接種之個體可為約1、約2、約3、約4、約5、約6、約7、約8、約9、約10、約11或約12個月齡。在一實施例中，待進行疫苗接種之個體為約2、約4或約6個月齡。在另一實施例中，待進行疫苗接種之個體小於2歲。舉例而言，待進行疫苗接種之個體可為約12個月齡至約15個月齡。在一些情況下，需要少至一次劑量之根據本發明之免疫原組合物，但在一些情況下，可給與第二次、第三次或第四次劑量(參見下方部分8)。In such embodiments, the individual to be vaccinated may be less than 1 year old. For example, the individual to be vaccinated may be about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, or about 12 months old. In one embodiment, the individual to be vaccinated is about 2, about 4, or about 6 months old. In another embodiment, the individual to be vaccinated is less than 2 years old. For example, the individual to be vaccinated may be about 12 months old to about 15 months old. In some cases, as little as one dose of the immunogenic composition according to the present invention is needed, but in some cases, a second, third, or fourth dose may be given (see Section 8 below).

在本發明之一實施例中，待進行疫苗接種之個體為50歲或更大之人類成人，更佳55歲或更大之人類成人。在一實施例中，待進行疫苗接種之個體為65歲或更大、70歲或更大、75歲或更大、或80歲或更大之人類成人。In one embodiment of the present invention, the individual to be vaccinated is a human adult of 50 years of age or older, more preferably a human adult of 55 years of age or older. In one embodiment, the individual to be vaccinated is a human adult of 65 years of age or older, 70 years of age or older, 75 years of age or older, or 80 years of age or older.

在一實施例中，待進行疫苗接種之個體為免疫功能不全個體，尤其人類。免疫功能不全個體一般定義為呈現建立對於感染劑刺激之正常體液或細胞防禦之能力減弱或降低的人。In one embodiment, the individual to be vaccinated is an immunocompromised individual, particularly a human. An immunocompromised individual is generally defined as a person who exhibits a weakened or reduced ability to mount a normal humoral or cellular defense against an infectious agent.

在本發明之一實施例中，待進行疫苗接種之免疫功能不全個體罹患削弱免疫系統且引起不足以防止肺炎鏈球菌疾病或治療肺炎鏈球菌疾病之抗體反應的疾病或病況。In one embodiment of the invention, the immunocompromised individual to be vaccinated suffers from a disease or condition that weakens the immune system and elicits an insufficient antibody response to prevent pneumococcal disease or to treat pneumococcal disease.

在一實施例中，該疾病為原發性免疫缺乏病症。較佳地，該原發性免疫缺乏病症係選自由以下組成之群：組合性T細胞及B細胞免疫缺乏症、抗體缺乏症、界定明確的症候群、免疫調節異常疾病、吞噬細胞病症、先天性免疫缺乏症、自體發炎性病症及補體缺乏症。在一實施例中，該原發性免疫缺乏病症係選自WO 2010/125480之第24頁第11行至第25頁第19行所揭露之病症。In one embodiment, the disease is a primary immunodeficiency disorder. Preferably, the primary immunodeficiency disorder is selected from the group consisting of combined T-cell and B-cell immunodeficiency, antibody deficiency, well-defined syndrome, abnormal immune regulation, phagocytic cell disease, congenital immune deficiency, autoinflammatory disease and complement deficiency. In one embodiment, the primary immunodeficiency disorder is selected from the disorders disclosed on page 24, line 11 to page 25, line 19 of WO 2010/125480.

在本發明之一特定實施例中，待進行疫苗接種之免疫功能不全個體罹患選自由以下組成之群之疾病：HIV感染、後天免疫缺乏症候群(AIDS)、癌症、慢性心臟病或肺病、充血性心臟衰竭、糖尿病、慢性肝病、酒精中毒、肝硬化、脊髓液滲漏、心肌病、慢性支氣管炎、肺氣腫、慢性阻塞性肺病(COPD)、脾功能障礙(諸如鐮狀細胞疾病)、脾功能缺乏(無脾症)、血液惡性疾病、白血病、多發性骨髓瘤、霍奇金氏疾病(Hodgkin's disease)、淋巴瘤、腎衰竭、腎病症候群及哮喘。In a specific embodiment of the present invention, the immunocompromised individual to be vaccinated suffers from a disease selected from the group consisting of HIV infection, acquired immune deficiency syndrome (AIDS), cancer, chronic heart disease or lung disease, congestive heart failure, diabetes, chronic liver disease, alcoholism, cirrhosis, cerebrospinal fluid leak, cardiomyopathy, chronic bronchitis, emphysema, chronic obstructive pulmonary disease (COPD), spleen dysfunction (such as sickle cell disease), spleen deficiency (asplenia), blood malignancies, leukemia, multiple myeloma, Hodgkin's disease, lymphoma, renal failure, nephrotic syndrome and asthma.

在本發明之一實施例中，待進行疫苗接種之免疫功能不全個體罹患營養不良。In one embodiment of the invention, the immunocompromised individual to be vaccinated suffers from malnutrition.

在本發明之一特定實施例中，待進行疫苗接種之免疫功能不全個體服用減少身體對於感染之抗性的藥物或治療。在一實施例中，該藥物係選自WO 2010/125480之第26頁第33行至第26頁第4行所揭露之藥物。In a specific embodiment of the present invention, the immunocompromised individual to be vaccinated takes a drug or treatment that reduces the body's resistance to infection. In one embodiment, the drug is selected from the drugs disclosed on page 26, line 33 to page 26, line 4 of WO 2010/125480.

在本發明之一特定實施例中，待進行疫苗接種之免疫功能不全個體為吸菸者。In a specific embodiment of the present invention, the immunocompromised individual to be vaccinated is a smoker.

在本發明之一特定實施例中，待進行疫苗接種之免疫功能不全個體具有以下之白血球計數(white blood cell count) (白血球計數(leukocyte count))：低於5 × 10 9個細胞/公升、或低於4 × 10 9個細胞/公升、或低於3 × 10 9個細胞/公升、或低於2 × 10 9個細胞/公升、或低於1 × 10 9個細胞/公升、或低於0.5 × 10 9個細胞/公升、或低於0.3 × 10 9個細胞/公升、或低於0.1 × 10 9個細胞/公升。 In a specific embodiment of the present invention, the immunocompromised individual to be vaccinated has a white blood cell count (leukocyte count): less than 5×10 9 cells/liter, or less than 4×10 9 cells/liter, or less than 3×10 9 cells/liter, or less than 2×10 9 cells/liter, or less than 1×10 9 cells/liter, or less than 0.5×10 9 cells/liter, or less than 0.3×10 9 cells/liter, or less than 0.1×10 9 cells/liter.

白血球計數(White blood cell count) (白血球計數(leukocyte count))：血液中之白血球(WBC)之數目。WBC通常作為CBC (全血球計數)之一部分量測。白血球為血液中之感染對抗細胞，且不同於被稱為紅血球之紅色(含氧)血細胞。存在不同類型的白血球，包括嗜中性球(多形核白血球；PMN)、帶狀細胞(略微不成熟的嗜中性球)、T型淋巴球(T細胞)、B型淋巴球(B細胞)、單核球、嗜酸性球及嗜鹼性球。所有類型之白血球均在白血球計數中反映。白血球計數之正常範圍通常在4,300與10,800個細胞/立方毫米血液之間。此亦可被稱作白細胞計數，且可以國際單位形式表述為4.3-10.8 × 10 9個細胞/公升。 White blood cell count (leukocyte count): The number of white blood cells (WBC) in the blood. WBC is usually measured as part of a CBC (complete blood count). Leukocytes are infection-fighting cells in the blood and are distinct from the red (oxygen-containing) blood cells, which are called erythrocytes. There are different types of white blood cells, including neutrophils (polymorphonuclear leukocytes; PMNs), band cells (slightly immature neutrophils), T-type lymphocytes (T cells), B-type lymphocytes (B cells), monocytes, eosinophils, and basophils. All types of white blood cells are reflected in the white blood cell count. The normal range for white blood cell count is usually between 4,300 and 10,800 cells/cubic millimeter of blood. This may also be called the white blood cell count and may be expressed in international units as 4.3-10.8 × 109 cells/L.

在本發明之一特定實施例中，待進行疫苗接種之免疫功能不全個體罹患嗜中性球減少症。在本發明之一特定實施例中，待進行疫苗接種之免疫功能不全個體具有以下之嗜中性球計數：低於2 × 10 9個細胞/公升、或低於1 × 10 9個細胞/公升、或低於0.5 × 10 9個細胞/公升、或低於0.1 × 10 9個細胞/公升、或低於0.05 × 10 9個細胞/公升。 In a specific embodiment of the invention, the immunocompromised individual to be vaccinated suffers from neutropenia. In a specific embodiment of the invention, the immunocompromised individual to be vaccinated has a neutrophil count of less than 2 × 10 9 cells/liter, or less than 1 × 10 9 cells/liter, or less than 0.5 × 10 9 cells/liter, or less than 0.1 × 10 9 cells/liter, or less than 0.05 × 10 9 cells/liter.

低白血球計數或「嗜中性球減少症」為特徵在於在循環血液中異常低含量之嗜中性球的病況。嗜中性球為幫助預防及對抗感染之一種特定種類之白血球。癌症患者經歷嗜中性球減少症之最常見原因為作為化學療法之副作用。化學療法誘發的嗜中性球減少症增加患者之感染風險且干擾癌症治療。Low white blood cell count or "neutropenia" is a condition characterized by abnormally low levels of neutrophils in the circulating blood. Neutrophils are a specific type of white blood cell that helps prevent and fight infection. The most common reason cancer patients experience neutropenia is as a side effect of chemotherapy. Chemotherapy-induced neutropenia increases a patient's risk of infection and interferes with cancer treatment.

在本發明之一特定實施例中，待進行疫苗接種之免疫功能不全個體具有低於500/mm 3之CD4+細胞計數、或低於300/mm 3之CD4+細胞計數、或低於200/mm 3之CD4+細胞計數、低於100/mm 3之CD4+細胞計數、低於75/mm 3之CD4+細胞計數、或低於50/mm 3之CD4+細胞計數。 In a specific embodiment of the invention, the immunocompromised individual to be vaccinated has a CD4+ cell count of less than 500/ mm3 , or a CD4+ cell count of less than 300/mm3, or a CD4+ cell count of less than 200/ mm3 , a CD4+ cell count of less than 100/ mm3 , a CD4+ cell count of less than 75/ mm3 , or a CD4+ cell count of less than 50/ mm3 .

CD4細胞測試通常報導為mm 3中之細胞數目。正常CD4計數在500與1,600之間，且CD8計數在375與1,100之間。CD4計數在患有HIV之人中急劇下降。 CD4 cell tests are usually reported as the number of cells in mm 3. Normal CD4 counts are between 500 and 1,600, and CD8 counts are between 375 and 1,100. CD4 counts drop dramatically in people with HIV.

在本發明之一實施例中，本文所揭露之免疫功能不全個體中之任一者為男性人類或女性人類。In one embodiment of the present invention, any of the immunocompromised individuals disclosed herein is a male human or a female human.

8 方案在一些情況下，需要少至一次劑量之根據本發明之免疫原組合物，但在一些情形下，諸如較高免疫缺乏之條件下，可給與第二次、第三次或第四次劑量。在初次疫苗接種之後，個體可接受一次或若干次恰當間隔的加強免疫接種。 8 regimen In some cases, as little as one dose of the immunogenic composition according to the invention is required, but in some cases, such as conditions of higher immunodeficiency, a second, third or fourth dose may be given. After the initial vaccination, individuals may receive one or several appropriately spaced booster vaccinations.

在一實施例中，根據本發明之免疫原組合物之疫苗接種之時程為單次劑量。在一特定實施例中，該單次劑量時程係針對至少2歲之健康人群。In one embodiment, the vaccination schedule of the immunogenic composition according to the present invention is a single dose. In a specific embodiment, the single dose schedule is for healthy people at least 2 years old.

在一實施例中，根據本發明之免疫原組合物之疫苗接種之時程為多劑量時程。在一特定實施例中，該多劑量時程由相隔約1個月至約2個月之時間間隔的一系列2次劑量組成。在一特定實施例中，該多劑量時程由相隔約1個月之時間間隔的一系列2次劑量組成，或由相隔約2個月之時間間隔的一系列2次劑量組成。In one embodiment, the vaccination schedule of the immunogenic composition according to the present invention is a multiple dose schedule. In a specific embodiment, the multiple dose schedule consists of a series of 2 doses separated by a time interval of about 1 month to about 2 months. In a specific embodiment, the multiple dose schedule consists of a series of 2 doses separated by a time interval of about 1 month, or consists of a series of 2 doses separated by a time interval of about 2 months.

在另一實施例中，該多劑量時程由相隔約1個月至約2個月之時間間隔的一系列3次劑量組成。在另一實施例中，該多劑量時程由相隔約1個月之時間間隔的一系列3次劑量組成，或由相隔約2個月之時間間隔的一系列3次劑量組成。In another embodiment, the multiple dose schedule consists of a series of 3 doses separated by a time interval of about 1 month to about 2 months. In another embodiment, the multiple dose schedule consists of a series of 3 doses separated by a time interval of about 1 month, or consists of a series of 3 doses separated by a time interval of about 2 months.

在另一實施例中，該多劑量時程由相隔約1個月至約2個月之時間間隔的一系列3次劑量及隨後在第一次劑量之後約10個月至約13個月之第四次劑量組成。在另一實施例中，該多劑量時程由相隔約1個月之時間間隔的一系列3次劑量及隨後在第一次劑量之後約10個月至約13個月之第四次劑量組成，或由相隔約2個月之時間間隔的一系列3次劑量及隨後在第一次劑量之後約10個月至約13個月之第四次劑量組成。In another embodiment, the multiple dose schedule consists of a series of 3 doses at intervals of about 1 month to about 2 months, followed by a fourth dose about 10 months to about 13 months after the first dose. In another embodiment, the multiple dose schedule consists of a series of 3 doses at intervals of about 1 month, followed by a fourth dose about 10 months to about 13 months after the first dose, or a series of 3 doses at intervals of about 2 months, followed by a fourth dose about 10 months to about 13 months after the first dose.

在一實施例中，多劑量時程由在一歲中之至少一次劑量(例如1、2或3次劑量)及隨後至少一次幼兒劑量組成。In one embodiment, the multiple-dose schedule consists of at least one dose (e.g., 1, 2, or 3 doses) in the first year of life followed by at least one pediatric dose.

在一實施例中，多劑量時程由以下組成：自2個月齡開始相隔約1個月至約2個月之時間間隔的一系列2或3次劑量(例如劑量之間相隔28至56天)，及隨後在12至18個月齡時之幼兒劑量。在一實施例中，該多劑量時程由以下組成：自2個月齡開始相隔約1個月至約2個月之時間間隔的一系列3次劑量(例如劑量之間相隔28至56天)，及隨後在12至15個月齡時之幼兒劑量。在另一實施例中，該多劑量時程由以下組成：自2個月齡開始相隔約2個月之時間間隔的一系列2次劑量，及隨後在12至18個月齡時之幼兒劑量。In one embodiment, the multiple-dose schedule consists of a series of 2 or 3 doses spaced about 1 month to about 2 months apart (e.g., 28 to 56 days between doses) beginning at 2 months of age, followed by a toddler dose at 12 to 18 months of age. In one embodiment, the multiple-dose schedule consists of a series of 3 doses spaced about 1 month to about 2 months apart (e.g., 28 to 56 days between doses) beginning at 2 months of age, followed by a toddler dose at 12 to 15 months of age. In another embodiment, the multiple-dose schedule consists of a series of 2 doses spaced about 2 months apart beginning at 2 months of age, followed by a pediatric dose at 12 to 18 months of age.

在一實施例中，該多劑量時程由在2、4、6及12至15個月齡時之4次劑量系列疫苗組成。In one embodiment, the multiple-dose schedule consists of a 4-dose series of vaccine at 2, 4, 6, and 12 to 15 months of age.

在一實施例中，在第0天給與初次劑量，且以在約2至約24週範圍內之間隔，較佳地以4至8週之給藥間隔，給與一或多次加強劑量。In one embodiment, a priming dose is administered on day 0, and one or more booster doses are administered at intervals ranging from about 2 to about 24 weeks, preferably at dosing intervals of 4 to 8 weeks.

在一實施例中，在第0天給與初次劑量，且在約3個月後給與加強劑量。In one embodiment, an initial dose is given on day 0 and a booster dose is given about 3 months later.

9. 分析方法在一實施例中，本發明係關於一種偵測經分離之肺炎鏈球菌血清型38多醣中之D-2-乙醯胺基-2,6-二去氧基-木糖-己-4-酮糖(D-Sug)殘基之存在的方法，該方法包含以下步驟：a)分離肺炎鏈球菌血清型38多醣及b)偵測該多醣中D-2-乙醯胺基-2,6-二去氧基-木糖-己-4-酮糖殘基之存在。 9. Analytical Methods In one embodiment, the present invention relates to a method for detecting the presence of D-2-acetamido-2,6-dideoxy-xylose-hex-4-ketose (D-Sug) residues in isolated Streptococcus pneumoniae serotype 38 polysaccharides, the method comprising the steps of: a) isolating Streptococcus pneumoniae serotype 38 polysaccharides and b) detecting the presence of D-2-acetamido-2,6-dideoxy-xylose-hex-4-ketose residues in the polysaccharide.

在一實施例中，藉由NMR或質譜法(MS)偵測到D-Sug殘基之存在。在一實施例中，藉由NMR偵測到D-Sug殘基之存在。在一實施例中，藉由1D NMR偵測到D-Sug殘基之存在。在一實施例中，藉由1D 1H或1D 13C NMR偵測到D-Sug殘基之存在。在一實施例中，藉由2D NMR偵測到D-Sug殘基之存在。在一實施例中，藉由異核單量子相干光譜法(HSQC)、異核多鍵相關光譜法(HMBC)、核奧氏效應光譜法(NOESY)、相關光譜法(COSY)、總相關光譜法(TOCSY)或異核單量子相干光譜法-總相關光譜法(HSQC-TOCSY)偵測到D-Sug殘基之存在。 In one embodiment, the presence of the D-Sug residue is detected by NMR or mass spectrometry (MS). In one embodiment, the presence of the D-Sug residue is detected by NMR. In one embodiment, the presence of the D-Sug residue is detected by 1D NMR. In one embodiment, the presence of the D-Sug residue is detected by 1D 1 H or 1D 13 C NMR. In one embodiment, the presence of the D-Sug residue is detected by 2D NMR. In one embodiment, the presence of the D-Sug residue is detected by heteronuclear single quantum coherence spectroscopy (HSQC), heteronuclear multibond correlation spectroscopy (HMBC), nuclear Ostwald effect spectroscopy (NOESY), correlation spectroscopy (COSY), total correlation spectroscopy (TOCSY), or heteronuclear single quantum coherence spectroscopy-total correlation spectroscopy (HSQC-TOCSY).

在一實施例中，藉由1D 1H、2D 1H- 13C異核單量子相干光譜法(HSQC)、2D 1H- 13C異核多鍵相關光譜法(HMBC)、2D 1H- 13C核奧氏效應光譜法(NOESY)、2D 1H- 13C相關光譜法(COSY)、2D 1H- 13C總相關光譜法(TOCSY)、2D 1H- 13C異核單量子相干光譜法-總相關光譜法(HSQC-TOCSY)或1D 13C NMR偵測到D-Sug殘基之存在。 In one embodiment, the presence of the D-Sug residue is detected by 1D 1 H, 2D 1 H- 13 C heteronuclear single quantum coherence spectroscopy (HSQC), 2D 1 H- 13 C heteronuclear multiple bond correlation spectroscopy (HMBC), 2D 1 H- 13 C nuclear Ostwald effect spectroscopy (NOESY), 2D 1 H- 13 C correlation spectroscopy (COSY), 2D 1 H- 13 C total correlation spectroscopy (TOCSY), 2D 1 H- 13 C heteronuclear single quantum coherence spectroscopy-total correlation spectroscopy (HSQC-TOCSY), or 1D 13 C NMR.

在一較佳實施例中，藉由1D 1H、2D 1H- 13C異核單量子相干光譜法(HSQC)或1D 13C NMR偵測到D-Sug殘基之存在。 In a preferred embodiment, the presence of the D-Sug residue is detected by 1D 1 H, 2D 1 H- 13 C heteronuclear single quantum coherence spectroscopy (HSQC) or 1D 13 C NMR.

在一實施例中，藉由2D 1H- 13C異核單量子相干光譜法(HSQC)偵測到D-Sug殘基之存在。 In one embodiment, the presence of the D-Sug residue is detected by 2D 1 H- 13 C heteronuclear single quantum coherence spectroscopy (HSQC).

在一實施例中，藉由質譜法(MS)偵測到D-Sug殘基之存在。在一實施例中，藉由串聯質譜法(MS/MS)偵測到D-Sug殘基之存在。在一實施例中，藉由氣相層析-質譜法(GC-MS)、液相層析-質譜法(LC-MS)、毛細管電泳-質譜法(CE-MS)或離子遷移譜-質譜法(IMS/MS或IMMS)偵測到D-Sug殘基之存在。在一實施例中，藉由尺寸排阻層析結合質譜法(SEC/MS)偵測到D-Sug殘基之存在。In one embodiment, the presence of D-Sug residues is detected by mass spectrometry (MS). In one embodiment, the presence of D-Sug residues is detected by tandem mass spectrometry (MS/MS). In one embodiment, the presence of D-Sug residues is detected by gas chromatography-mass spectrometry (GC-MS), liquid chromatography-mass spectrometry (LC-MS), capillary electrophoresis-mass spectrometry (CE-MS), or ion mobility shift spectrometry-mass spectrometry (IMS/MS or IMMS). In one embodiment, the presence of D-Sug residues is detected by size exclusion chromatography combined with mass spectrometry (SEC/MS).

在一實施例中，藉由氣相層析-質譜法(GC-MS)偵測到D-Sug殘基之存在。在一實施例中，藉由液相層析-質譜法(LC-MS)偵測到D-Sug殘基之存在。在一實施例中，藉由毛細管電泳-質譜法(CE-MS)偵測到D-Sug殘基之存在。在一實施例中，藉由離子遷移譜-質譜法(IMS/MS)偵測到D-Sug殘基之存在。在一實施例中，藉由親水相互作用液相層析-質譜法(HILIC-LC/MS)偵測到D-Sug殘基之存在。In one embodiment, the presence of D-Sug residues is detected by gas chromatography-mass spectrometry (GC-MS). In one embodiment, the presence of D-Sug residues is detected by liquid chromatography-mass spectrometry (LC-MS). In one embodiment, the presence of D-Sug residues is detected by capillary electrophoresis-mass spectrometry (CE-MS). In one embodiment, the presence of D-Sug residues is detected by ion mobility shift spectrometry-mass spectrometry (IMS/MS). In one embodiment, the presence of D-Sug residues is detected by hydrophilic interaction liquid chromatography-mass spectrometry (HILIC-LC/MS).

在一實施例中，本發明係關於一種偵測經還原之血清型38多醣中之N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在的方法，該方法包含以下步驟：a)使經分離之肺炎鏈球菌血清型38多醣與還原劑反應，及b)偵測該經還原之多醣中N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。In one embodiment, the present invention relates to a method for detecting the presence of N-acetyl-D-fucosamine (D-FucNAc) residues in reduced serotype 38 polysaccharides, the method comprising the steps of: a) reacting isolated Streptococcus pneumoniae serotype 38 polysaccharides with a reducing agent, and b) detecting the presence of N-acetyl-D-fucosamine (D-FucNAc) residues in the reduced polysaccharide.

在一實施例中，藉由NMR偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。在一實施例中，藉由2D NMR偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) residues is detected by NMR. In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) residues is detected by 2D NMR.

在一較佳實施例中，藉由2D 1H- 13C HSQC NMR偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。 In a preferred embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) residue is detected by 2D 1 H- 13 C HSQC NMR.

在一實施例中，藉由質譜法(MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。在一實施例中，藉由串聯質譜法(MS/MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。在一實施例中，藉由氣相層析-質譜法(GC-MS)、液相層析-質譜法(LC-MS)、毛細管電泳-質譜法(CE-MS)或離子遷移譜-質譜法(IMS/MS或IMMS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。在一實施例中，藉由尺寸排阻層析結合質譜法(SEC/MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) residues is detected by mass spectrometry (MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) residues is detected by tandem mass spectrometry (MS/MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) residues is detected by gas chromatography-mass spectrometry (GC-MS), liquid chromatography-mass spectrometry (LC-MS), capillary electrophoresis-mass spectrometry (CE-MS), or ion mobility shift spectrometry-mass spectrometry (IMS/MS or IMMS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) residues is detected by size exclusion chromatography coupled to mass spectrometry (SEC/MS).

在一實施例中，藉由氣相層析-質譜法(GC-MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。在一實施例中，藉由液相層析-質譜法(LC-MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。在一實施例中，藉由毛細管電泳-質譜法(CE-MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。在一實施例中，藉由離子遷移譜-質譜法(IMS/MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。在一實施例中，藉由親水相互作用液相層析-質譜法(HILIC-LC/MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) residues is detected by gas chromatography-mass spectrometry (GC-MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) residues is detected by liquid chromatography-mass spectrometry (LC-MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) residues is detected by capillary electrophoresis-mass spectrometry (CE-MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) residues is detected by ion mobility shift spectrometry-mass spectrometry (IMS/MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) residues is detected by hydrophilic interaction liquid chromatography-mass spectrometry (HILIC-LC/MS).

在一實施例中，該還原劑為硼氫化鈉(NaBH 4)。 In one embodiment, the reducing agent is sodium borohydride (NaBH 4 ).

在一實施例中，該經分離之肺炎鏈球菌血清型38多醣先前已用氧化劑處理。在一實施例中，氧化劑為將末端羥基氧化成醛之任何氧化劑。在一實施例中，氧化劑為過碘酸鹽。在一實施例中，氧化劑為正過碘酸鹽。在一個較佳實施例中，氧化劑為過碘酸鈉。在一實施例中，氧化劑為偏過碘酸鹽。在一較佳實施例中，氧化劑為偏過碘酸鈉。In one embodiment, the isolated S. pneumoniae serotype 38 polysaccharide has been previously treated with an oxidizing agent. In one embodiment, the oxidizing agent is any oxidizing agent that oxidizes a terminal hydroxyl group to an aldehyde. In one embodiment, the oxidizing agent is a periodate. In one embodiment, the oxidizing agent is orthoperiodate. In a preferred embodiment, the oxidizing agent is sodium periodate. In one embodiment, the oxidizing agent is metaperiodate. In a preferred embodiment, the oxidizing agent is sodium metaperiodate.

在一實施例中，該經分離之肺炎鏈球菌血清型38多醣先前已用穩定硝醯基自由基化合物及氧化劑處理。在一態樣中，該穩定硝醯基自由基化合物為帶有TEMPO或PROXYL (2,2,5,5-四甲基-1-吡咯啶基氧基)部分之分子。較佳地，該分子能夠在氧化劑存在下選擇性地氧化一級醇，以產生醛基，而不影響二級羥基。更佳地，該分子能夠在氧化劑存在下選擇性地氧化一級醇，以產生醛基，而不過度氧化成羧基。在一態樣中，該穩定硝醯基自由基化合物為TEMPO、2,2,6,6-四甲基-4-(甲基磺醯氧基)-1-哌啶氧基、4-膦醯氧基-TEMPO、4-側氧基-TEMPO、4-甲氧基-TEMPO、4-異硫氰基-TEMPO、4-(2-碘乙醯胺基)-TEMPO自由基、4-羥基-TEMPO、4-氰基-TEMPO、4-羧基-TEMPO、4-(2-溴乙醯胺基)-TEMPO、4-胺基-TEMPO或4-乙醯胺基-2,2,6,6-四甲基哌啶1-氧基。在一態樣中，該穩定硝醯基自由基化合物係選自由以下組成之群：TEMPO、2,2,6,6-四甲基-4-(甲基磺醯氧基)-1-哌啶氧基、4-膦醯氧基-TEMPO、4-側氧基-TEMPO、4-甲氧基-TEMPO、4-異硫氰基-TEMPO、4-(2-碘乙醯胺基)-TEMPO自由基、4-羥基-TEMPO、4-氰基-TEMPO、4-羧基-TEMPO、4-(2-溴乙醯胺基)-TEMPO、4-胺基-TEMPO、4-乙醯胺基-2,2,6,6-四甲基哌啶1-氧基。較佳地，該穩定硝醯基自由基化合物為TEMPO。在另一態樣中，該穩定硝醯基自由基化合物為3β-DOXYL-5α-膽甾烷、5-DOXYL-硬脂酸、16-DOXYL-硬脂酸、5-DOXYL-硬脂酸甲酯、3-(胺基甲基)-PROXYL、3-胺甲醯基-PROXYL、3-胺甲醯基-2,2,5,5-四甲基-3-吡咯啉-1-氧基、3-羧基-PROXYL或3-氰基-PROXYL。在另一態樣中，該穩定硝醯基自由基化合物係選自由以下組成之群：3β-DOXYL-5α-膽甾烷、5-DOXYL-硬脂酸、16-DOXYL-硬脂酸、5-DOXYL-硬脂酸甲酯、3-(胺基甲基)-PROXYL、3-胺甲醯基-PROXYL、3-胺甲醯基-2,2,5,5-四甲基-3-吡咯啉-1-氧基、3-羧基-PROXYL、3-氰基-PROXYL。在一態樣中，氧化劑為帶有N-鹵基部分之分子。較佳地，該分子能夠在硝醯基自由基化合物存在下選擇性氧化一級醇。在一態樣中，該氧化劑為N-氯代丁二醯亞胺、N-溴代丁二醯亞胺、N-碘丁二醯亞胺、二氯異三聚氰酸、1,3,5-三氯-1,3,5-三𠯤烷-2,4,6-三酮、二溴異三聚氰酸、1,3,5-三溴-1,3,5-三𠯤烷-2,4,6-三酮、二碘異三聚氰酸或1,3,5-三碘-1,3,5-三𠯤烷-2,4,6-三酮。在一態樣中，該氧化劑係選自由以下組成之群：N-氯代丁二醯亞胺、N-溴代丁二醯亞胺、N-碘丁二醯亞胺、二氯異三聚氰酸、1,3,5-三氯-1,3,5-三𠯤烷-2,4,6-三酮、二溴異三聚氰酸、1,3,5-三溴-1,3,5-三𠯤烷-2,4,6-三酮、二碘異三聚氰酸及1,3,5-三碘-1,3,5-三𠯤烷-2,4,6-三酮。較佳地，該氧化劑為N-氯代丁二醯亞胺。In one embodiment, the isolated S. pneumoniae serotype 38 polysaccharide has been previously treated with a stable nitro radical compound and an oxidizing agent. In one aspect, the stable nitro radical compound is a molecule with a TEMPO or PROXYL (2,2,5,5-tetramethyl-1-pyrrolidinyloxy) moiety. Preferably, the molecule is capable of selectively oxidizing a primary alcohol in the presence of an oxidizing agent to produce an aldehyde group without affecting a secondary hydroxyl group. More preferably, the molecule is capable of selectively oxidizing a primary alcohol in the presence of an oxidizing agent to produce an aldehyde group without excessive oxidation to a carboxyl group. In one embodiment, the stable nitro radical compound is TEMPO, 2,2,6,6-tetramethyl-4-(methylsulfonyloxy)-1-piperidinyloxy, 4-phosphonyloxy-TEMPO, 4-oxo-TEMPO, 4-methoxy-TEMPO, 4-isothiocyanato-TEMPO, 4-(2-iodoacetamido)-TEMPO radical, 4-hydroxy-TEMPO, 4-cyano-TEMPO, 4-carboxy-TEMPO, 4-(2-bromoacetamido)-TEMPO, 4-amino-TEMPO or 4-acetamido-2,2,6,6-tetramethylpiperidinyl-1-oxyl. In one embodiment, the stable nitro radical compound is selected from the group consisting of TEMPO, 2,2,6,6-tetramethyl-4-(methylsulfonyloxy)-1-piperidinyloxy, 4-phosphonyloxy-TEMPO, 4-oxo-TEMPO, 4-methoxy-TEMPO, 4-isothiocyanato-TEMPO, 4-(2-iodoacetamido)-TEMPO radical, 4-hydroxy-TEMPO, 4-cyano-TEMPO, 4-carboxyl-TEMPO, 4-(2-bromoacetamido)-TEMPO, 4-amino-TEMPO, 4-acetamido-2,2,6,6-tetramethylpiperidinyl-1-oxyl. Preferably, the stable nitro radical compound is TEMPO. In another embodiment, the stable nitro radical compound is 3β-DOXYL-5α-cholestane, 5-DOXYL-stearic acid, 16-DOXYL-stearic acid, 5-DOXYL-stearate methyl ester, 3-(aminomethyl)-PROXYL, 3-aminoformyl-PROXYL, 3-aminoformyl-2,2,5,5-tetramethyl-3-pyrroline-1-oxyl, 3-carboxy-PROXYL or 3-cyano-PROXYL. In another embodiment, the stable nitro radical compound is selected from the group consisting of: 3β-DOXYL-5α-cholestane, 5-DOXYL-stearic acid, 16-DOXYL-stearic acid, 5-DOXYL-stearate methyl ester, 3-(aminomethyl)-PROXYL, 3-aminoformyl-PROXYL, 3-aminoformyl-2,2,5,5-tetramethyl-3-pyrroline-1-oxyl, 3-carboxyl-PROXYL, 3-cyano-PROXYL. In one embodiment, the oxidizing agent is a molecule with an N-halogen moiety. Preferably, the molecule is capable of selectively oxidizing a primary alcohol in the presence of a nitro radical compound. In one embodiment, the oxidizing agent is N-chlorosuccinimide, N-bromosuccinimide, N-iodosuccinimide, dichloroisocyanuric acid, 1,3,5-trichloro-1,3,5-trioxane-2,4,6-trione, dibromoisocyanuric acid, 1,3,5-tribromo-1,3,5-trioxane-2,4,6-trione, diiodoisocyanuric acid or 1,3,5-triiodo-1,3,5-trioxane-2,4,6-trione. In one embodiment, the oxidant is selected from the group consisting of: N-chlorosuccinimide, N-bromosuccinimide, N-iodosuccinimide, dichloroisocyanuric acid, 1,3,5-trichloro-1,3,5-trioxane-2,4,6-trione, dibromoisocyanuric acid, 1,3,5-tribromo-1,3,5-trioxane-2,4,6-trione, diiodoisocyanuric acid and 1,3,5-triiodo-1,3,5-trioxane-2,4,6-trione. Preferably, the oxidant is N-chlorosuccinimide.

在一態樣中，該穩定硝醯基自由基化合物為2,2,6,6-四甲基-1-哌啶基氧基自由基(TEMPO)，且該氧化劑為N-氯代丁二醯亞胺(NCS)。In one embodiment, the stable nitroxyl radical compound is 2,2,6,6-tetramethyl-1-piperidinyloxy radical (TEMPO), and the oxidizing agent is N-chlorosuccinimide (NCS).

在一實施例中，本發明係關於一種偵測經還原之血清型38多醣中之N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在的方法，該方法包含以下步驟：a)使經分離之肺炎鏈球菌血清型38多醣與還原劑反應，及b)偵測該經還原之多醣中N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。In one embodiment, the present invention relates to a method for detecting the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues in reduced serotype 38 polysaccharides, the method comprising the steps of: a) reacting isolated Streptococcus pneumoniae serotype 38 polysaccharides with a reducing agent, and b) detecting the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues in the reduced polysaccharide.

在一實施例中，藉由NMR偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由2D NMR偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。In one embodiment, the presence of the N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue is detected by NMR. In one embodiment, the presence of the N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue is detected by 2D NMR.

在一實施例中，藉由異核單量子相干光譜法(HSQC)、異核多鍵相關光譜法(HMBC)、相關光譜法(COSY)及/或異核單量子相干光譜法-總相關光譜法(HSQC-TOCSY)偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。In one embodiment, the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue is detected by heteronuclear single quantum coherence spectroscopy (HSQC), heteronuclear multibond correlation spectroscopy (HMBC), correlation spectroscopy (COSY) and/or heteronuclear single quantum coherence spectroscopy-total correlation spectroscopy (HSQC-TOCSY).

在一較佳實施例中，藉由2D 1H- 13C HSQC NMR偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。 In a preferred embodiment, the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue is detected by 2D 1 H- 13 C HSQC NMR.

在一實施例中，藉由質譜法(MS)偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由串聯質譜法(MS/MS)偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由氣相層析-質譜法(GC-MS)、液相層析-質譜法(LC-MS)、毛細管電泳-質譜法(CE-MS)或離子遷移譜-質譜法(IMS/MS或IMMS)偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由尺寸排阻層析結合質譜法(SEC/MS)偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。In one embodiment, the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue is detected by mass spectrometry (MS). In one embodiment, the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue is detected by tandem mass spectrometry (MS/MS). In one embodiment, the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue is detected by gas chromatography-mass spectrometry (GC-MS), liquid chromatography-mass spectrometry (LC-MS), capillary electrophoresis-mass spectrometry (CE-MS) or ion mobility shift spectrometry-mass spectrometry (IMS/MS or IMMS). In one embodiment, the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue is detected by size exclusion chromatography coupled to mass spectrometry (SEC/MS).

在一實施例中，藉由氣相層析-質譜法(GC-MS)偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由液相層析-質譜法(LC-MS)偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由毛細管電泳-質譜法(CE-MS)偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由離子遷移譜-質譜法(IMS/MS)偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由親水相互作用液相層析-質譜法(HILIC-LC/MS)偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。In one embodiment, the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by gas chromatography-mass spectrometry (GC-MS). In one embodiment, the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by liquid chromatography-mass spectrometry (LC-MS). In one embodiment, the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by capillary electrophoresis-mass spectrometry (CE-MS). In one embodiment, the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by ion mobility shift spectrometry-mass spectrometry (IMS/MS). In one embodiment, the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by hydrophilic interaction liquid chromatography-mass spectrometry (HILIC-LC/MS).

在一實施例中，該還原劑為硼氫化鈉(NaBH 4)。 In one embodiment, the reducing agent is sodium borohydride (NaBH 4 ).

在一實施例中，該經分離之肺炎鏈球菌血清型38多醣先前已用氧化劑處理。在一實施例中，氧化劑為將末端羥基氧化成醛之任何氧化劑。在一實施例中，氧化劑為過碘酸鹽。在一實施例中，氧化劑為正過碘酸鹽。在一個較佳實施例中，氧化劑為過碘酸鈉。在一實施例中，氧化劑為偏過碘酸鹽。在一較佳實施例中，氧化劑為偏過碘酸鈉。In one embodiment, the isolated S. pneumoniae serotype 38 polysaccharide has been previously treated with an oxidizing agent. In one embodiment, the oxidizing agent is any oxidizing agent that oxidizes a terminal hydroxyl group to an aldehyde. In one embodiment, the oxidizing agent is a periodate. In one embodiment, the oxidizing agent is orthoperiodate. In a preferred embodiment, the oxidizing agent is sodium periodate. In one embodiment, the oxidizing agent is metaperiodate. In a preferred embodiment, the oxidizing agent is sodium metaperiodate.

在一實施例中，該經分離之肺炎鏈球菌血清型38多醣先前已用穩定硝醯基自由基化合物及氧化劑處理。在一態樣中，該穩定硝醯基自由基化合物為帶有TEMPO或PROXYL (2,2,5,5-四甲基-1-吡咯啶基氧基)部分之分子。較佳地，該分子能夠在氧化劑存在下選擇性地氧化一級醇，以產生醛基，而不影響二級羥基。更佳地，該分子能夠在氧化劑存在下選擇性地氧化一級醇，以產生醛基，而不過度氧化成羧基。在一態樣中，該穩定硝醯基自由基化合物為TEMPO、2,2,6,6-四甲基-4-(甲基磺醯氧基)-1-哌啶氧基、4-膦醯氧基-TEMPO、4-側氧基-TEMPO、4-甲氧基-TEMPO、4-異硫氰基-TEMPO、4-(2-碘乙醯胺基)-TEMPO自由基、4-羥基-TEMPO、4-氰基-TEMPO、4-羧基-TEMPO、4-(2-溴乙醯胺基)-TEMPO、4-胺基-TEMPO或4-乙醯胺基-2,2,6,6-四甲基哌啶1-氧基。在一態樣中，該穩定硝醯基自由基化合物係選自由以下組成之群：TEMPO、2,2,6,6-四甲基-4-(甲基磺醯氧基)-1-哌啶氧基、4-膦醯氧基-TEMPO、4-側氧基-TEMPO、4-甲氧基-TEMPO、4-異硫氰基-TEMPO、4-(2-碘乙醯胺基)-TEMPO自由基、4-羥基-TEMPO、4-氰基-TEMPO、4-羧基-TEMPO、4-(2-溴乙醯胺基)-TEMPO、4-胺基-TEMPO、4-乙醯胺基-2,2,6,6-四甲基哌啶1-氧基。較佳地，該穩定硝醯基自由基化合物為TEMPO。在另一態樣中，該穩定硝醯基自由基化合物為3β-DOXYL-5α-膽甾烷、5-DOXYL-硬脂酸、16-DOXYL-硬脂酸、5-DOXYL-硬脂酸甲酯、3-(胺基甲基)-PROXYL、3-胺甲醯基-PROXYL、3-胺甲醯基-2,2,5,5-四甲基-3-吡咯啉-1-氧基、3-羧基-PROXYL或3-氰基-PROXYL。在另一態樣中，該穩定硝醯基自由基化合物係選自由以下組成之群：3β-DOXYL-5α-膽甾烷、5-DOXYL-硬脂酸、16-DOXYL-硬脂酸、5-DOXYL-硬脂酸甲酯、3-(胺基甲基)-PROXYL、3-胺甲醯基-PROXYL、3-胺甲醯基-2,2,5,5-四甲基-3-吡咯啉-1-氧基、3-羧基-PROXYL、3-氰基-PROXYL。在一態樣中，氧化劑為帶有N-鹵基部分之分子。較佳地，該分子能夠在硝醯基自由基化合物存在下選擇性氧化一級醇。在一態樣中，該氧化劑為N-氯代丁二醯亞胺、N-溴代丁二醯亞胺、N-碘丁二醯亞胺、二氯異三聚氰酸、1,3,5-三氯-1,3,5-三𠯤烷-2,4,6-三酮、二溴異三聚氰酸、1,3,5-三溴-1,3,5-三𠯤烷-2,4,6-三酮、二碘異三聚氰酸或1,3,5-三碘-1,3,5-三𠯤烷-2,4,6-三酮。在一態樣中，該氧化劑係選自由以下組成之群：N-氯代丁二醯亞胺、N-溴代丁二醯亞胺、N-碘丁二醯亞胺、二氯異三聚氰酸、1,3,5-三氯-1,3,5-三𠯤烷-2,4,6-三酮、二溴異三聚氰酸、1,3,5-三溴-1,3,5-三𠯤烷-2,4,6-三酮、二碘異三聚氰酸及1,3,5-三碘-1,3,5-三𠯤烷-2,4,6-三酮。較佳地，該氧化劑為N-氯代丁二醯亞胺。In one embodiment, the isolated S. pneumoniae serotype 38 polysaccharide has been previously treated with a stable nitro radical compound and an oxidizing agent. In one aspect, the stable nitro radical compound is a molecule with a TEMPO or PROXYL (2,2,5,5-tetramethyl-1-pyrrolidinyloxy) moiety. Preferably, the molecule is capable of selectively oxidizing a primary alcohol in the presence of an oxidizing agent to produce an aldehyde group without affecting a secondary hydroxyl group. More preferably, the molecule is capable of selectively oxidizing a primary alcohol in the presence of an oxidizing agent to produce an aldehyde group without excessive oxidation to a carboxyl group. In one embodiment, the stable nitro radical compound is TEMPO, 2,2,6,6-tetramethyl-4-(methylsulfonyloxy)-1-piperidinyloxy, 4-phosphonyloxy-TEMPO, 4-oxo-TEMPO, 4-methoxy-TEMPO, 4-isothiocyanato-TEMPO, 4-(2-iodoacetamido)-TEMPO radical, 4-hydroxy-TEMPO, 4-cyano-TEMPO, 4-carboxy-TEMPO, 4-(2-bromoacetamido)-TEMPO, 4-amino-TEMPO or 4-acetamido-2,2,6,6-tetramethylpiperidinyl-1-oxyl. In one embodiment, the stable nitro radical compound is selected from the group consisting of TEMPO, 2,2,6,6-tetramethyl-4-(methylsulfonyloxy)-1-piperidinyloxy, 4-phosphonyloxy-TEMPO, 4-oxo-TEMPO, 4-methoxy-TEMPO, 4-isothiocyanato-TEMPO, 4-(2-iodoacetamido)-TEMPO radical, 4-hydroxy-TEMPO, 4-cyano-TEMPO, 4-carboxyl-TEMPO, 4-(2-bromoacetamido)-TEMPO, 4-amino-TEMPO, 4-acetamido-2,2,6,6-tetramethylpiperidinyl-1-oxyl. Preferably, the stable nitro radical compound is TEMPO. In another embodiment, the stable nitro radical compound is 3β-DOXYL-5α-cholestane, 5-DOXYL-stearic acid, 16-DOXYL-stearic acid, 5-DOXYL-stearate methyl ester, 3-(aminomethyl)-PROXYL, 3-aminoformyl-PROXYL, 3-aminoformyl-2,2,5,5-tetramethyl-3-pyrroline-1-oxyl, 3-carboxy-PROXYL or 3-cyano-PROXYL. In another embodiment, the stable nitro radical compound is selected from the group consisting of: 3β-DOXYL-5α-cholestane, 5-DOXYL-stearic acid, 16-DOXYL-stearic acid, 5-DOXYL-stearate methyl ester, 3-(aminomethyl)-PROXYL, 3-aminoformyl-PROXYL, 3-aminoformyl-2,2,5,5-tetramethyl-3-pyrroline-1-oxyl, 3-carboxyl-PROXYL, 3-cyano-PROXYL. In one embodiment, the oxidizing agent is a molecule with an N-halogen moiety. Preferably, the molecule is capable of selectively oxidizing a primary alcohol in the presence of a nitro radical compound. In one embodiment, the oxidizing agent is N-chlorosuccinimide, N-bromosuccinimide, N-iodosuccinimide, dichloroisocyanuric acid, 1,3,5-trichloro-1,3,5-trioxane-2,4,6-trione, dibromoisocyanuric acid, 1,3,5-tribromo-1,3,5-trioxane-2,4,6-trione, diiodoisocyanuric acid or 1,3,5-triiodo-1,3,5-trioxane-2,4,6-trione. In one embodiment, the oxidant is selected from the group consisting of: N-chlorosuccinimide, N-bromosuccinimide, N-iodosuccinimide, dichloroisocyanuric acid, 1,3,5-trichloro-1,3,5-trioxane-2,4,6-trione, dibromoisocyanuric acid, 1,3,5-tribromo-1,3,5-trioxane-2,4,6-trione, diiodoisocyanuric acid and 1,3,5-triiodo-1,3,5-trioxane-2,4,6-trione. Preferably, the oxidant is N-chlorosuccinimide.

在一態樣中，該穩定硝醯基自由基化合物為2,2,6,6-四甲基-1-哌啶基氧基自由基(TEMPO)，且該氧化劑為N-氯代丁二醯亞胺(NCS)。In one embodiment, the stable nitroxyl radical compound is 2,2,6,6-tetramethyl-1-piperidinyloxy radical (TEMPO), and the oxidizing agent is N-chlorosuccinimide (NCS).

在一實施例中，本發明係關於一種偵測經還原之血清型38多醣中之N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在的方法，該方法包含以下步驟：a)使經分離之肺炎鏈球菌血清型38多醣與還原劑反應，及b)偵測該經還原之多醣中N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。In one embodiment, the present invention relates to a method for detecting the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfuramine (D-QuiNAc) residues in reduced serotype 38 polysaccharides, the method comprising the steps of: a) reacting isolated Streptococcus pneumoniae serotype 38 polysaccharides with a reducing agent, and b) detecting the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfuramine (D-QuiNAc) residues in the reduced polysaccharide.

在一實施例中，藉由NMR偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by NMR.

在一較佳實施例中，藉由2D-NMR偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。In a preferred embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by 2D-NMR.

在一實施例中，藉由質譜法(MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由串聯質譜法(MS/MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由氣相層析-質譜法(GC-MS)、液相層析-質譜法(LC-MS)、毛細管電泳-質譜法(CE-MS)或離子遷移譜-質譜法(IMS/MS或IMMS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由尺寸排阻層析結合質譜法(SEC/MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by mass spectrometry (MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by tandem mass spectrometry (MS/MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by gas chromatography-mass spectrometry (GC-MS), liquid chromatography-mass spectrometry (LC-MS), capillary electrophoresis-mass spectrometry (CE-MS), or ion mobility shift spectrometry-mass spectrometry (IMS/MS or IMMS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by size exclusion chromatography combined with mass spectrometry (SEC/MS).

在一實施例中，藉由氣相層析-質譜法(GC-MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由液相層析-質譜法(LC-MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由毛細管電泳-質譜法(CE-MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由離子遷移譜-質譜法(IMS/MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由親水相互作用液相層析-質譜法(HILIC-LC/MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by gas chromatography-mass spectrometry (GC-MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by liquid chromatography-mass spectrometry (LC-MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by capillary electrophoresis-mass spectrometry (CE-MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by ion mobility shift spectrometry-mass spectrometry (IMS/MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by hydrophilic interaction liquid chromatography-mass spectrometry (HILIC-LC/MS).

在一實施例中，該還原劑為硼氫化鈉(NaBH 4)。 In one embodiment, the reducing agent is sodium borohydride (NaBH 4 ).

在一實施例中，該經分離之肺炎鏈球菌血清型38多醣先前已用氧化劑處理。在一實施例中，氧化劑為將末端羥基氧化成醛之任何氧化劑。在一實施例中，氧化劑為過碘酸鹽。在一實施例中，氧化劑為正過碘酸鹽。在一個較佳實施例中，氧化劑為過碘酸鈉。在一實施例中，氧化劑為偏過碘酸鹽。在一較佳實施例中，氧化劑為偏過碘酸鈉。In one embodiment, the isolated S. pneumoniae serotype 38 polysaccharide has been previously treated with an oxidizing agent. In one embodiment, the oxidizing agent is any oxidizing agent that oxidizes a terminal hydroxyl group to an aldehyde. In one embodiment, the oxidizing agent is a periodate. In one embodiment, the oxidizing agent is orthoperiodate. In a preferred embodiment, the oxidizing agent is sodium periodate. In one embodiment, the oxidizing agent is metaperiodate. In a preferred embodiment, the oxidizing agent is sodium metaperiodate.

在一實施例中，該經分離之肺炎鏈球菌血清型38多醣先前已用穩定硝醯基自由基化合物及氧化劑處理。在一態樣中，該穩定硝醯基自由基化合物為帶有TEMPO或PROXYL (2,2,5,5-四甲基-1-吡咯啶基氧基)部分之分子。較佳地，該分子能夠在氧化劑存在下選擇性地氧化一級醇，以產生醛基，而不影響二級羥基。更佳地，該分子能夠在氧化劑存在下選擇性地氧化一級醇，以產生醛基，而不過度氧化成羧基。在一態樣中，該穩定硝醯基自由基化合物為TEMPO、2,2,6,6-四甲基-4-(甲基磺醯氧基)-1-哌啶氧基、4-膦醯氧基-TEMPO、4-側氧基-TEMPO、4-甲氧基-TEMPO、4-異硫氰基-TEMPO、4-(2-碘乙醯胺基)-TEMPO自由基、4-羥基-TEMPO、4-氰基-TEMPO、4-羧基-TEMPO、4-(2-溴乙醯胺基)-TEMPO、4-胺基-TEMPO或4-乙醯胺基-2,2,6,6-四甲基哌啶1-氧基。在一態樣中，該穩定硝醯基自由基化合物係選自由以下組成之群：TEMPO、2,2,6,6-四甲基-4-(甲基磺醯氧基)-1-哌啶氧基、4-膦醯氧基-TEMPO、4-側氧基-TEMPO、4-甲氧基-TEMPO、4-異硫氰基-TEMPO、4-(2-碘乙醯胺基)-TEMPO自由基、4-羥基-TEMPO、4-氰基-TEMPO、4-羧基-TEMPO、4-(2-溴乙醯胺基)-TEMPO、4-胺基-TEMPO、4-乙醯胺基-2,2,6,6-四甲基哌啶1-氧基。較佳地，該穩定硝醯基自由基化合物為TEMPO。在另一態樣中，該穩定硝醯基自由基化合物為3β-DOXYL-5α-膽甾烷、5-DOXYL-硬脂酸、16-DOXYL-硬脂酸、5-DOXYL-硬脂酸甲酯、3-(胺基甲基)-PROXYL、3-胺甲醯基-PROXYL、3-胺甲醯基-2,2,5,5-四甲基-3-吡咯啉-1-氧基、3-羧基-PROXYL或3-氰基-PROXYL。在另一態樣中，該穩定硝醯基自由基化合物係選自由以下組成之群：3β-DOXYL-5α-膽甾烷、5-DOXYL-硬脂酸、16-DOXYL-硬脂酸、5-DOXYL-硬脂酸甲酯、3-(胺基甲基)-PROXYL、3-胺甲醯基-PROXYL、3-胺甲醯基-2,2,5,5-四甲基-3-吡咯啉-1-氧基、3-羧基-PROXYL、3-氰基-PROXYL。在一態樣中，氧化劑為帶有N-鹵基部分之分子。較佳地，該分子能夠在硝醯基自由基化合物存在下選擇性氧化一級醇。在一態樣中，該氧化劑為N-氯代丁二醯亞胺、N-溴代丁二醯亞胺、N-碘丁二醯亞胺、二氯異三聚氰酸、1,3,5-三氯-1,3,5-三𠯤烷-2,4,6-三酮、二溴異三聚氰酸、1,3,5-三溴-1,3,5-三𠯤烷-2,4,6-三酮、二碘異三聚氰酸或1,3,5-三碘-1,3,5-三𠯤烷-2,4,6-三酮。在一態樣中，該氧化劑係選自由以下組成之群：N-氯代丁二醯亞胺、N-溴代丁二醯亞胺、N-碘丁二醯亞胺、二氯異三聚氰酸、1,3,5-三氯-1,3,5-三𠯤烷-2,4,6-三酮、二溴異三聚氰酸、1,3,5-三溴-1,3,5-三𠯤烷-2,4,6-三酮、二碘異三聚氰酸及1,3,5-三碘-1,3,5-三𠯤烷-2,4,6-三酮。較佳地，該氧化劑為N-氯代丁二醯亞胺。In one embodiment, the isolated S. pneumoniae serotype 38 polysaccharide has been previously treated with a stable nitro radical compound and an oxidizing agent. In one aspect, the stable nitro radical compound is a molecule with a TEMPO or PROXYL (2,2,5,5-tetramethyl-1-pyrrolidinyloxy) moiety. Preferably, the molecule is capable of selectively oxidizing a primary alcohol in the presence of an oxidizing agent to produce an aldehyde group without affecting a secondary hydroxyl group. More preferably, the molecule is capable of selectively oxidizing a primary alcohol in the presence of an oxidizing agent to produce an aldehyde group without excessive oxidation to a carboxyl group. In one embodiment, the stable nitro radical compound is TEMPO, 2,2,6,6-tetramethyl-4-(methylsulfonyloxy)-1-piperidinyloxy, 4-phosphonyloxy-TEMPO, 4-oxo-TEMPO, 4-methoxy-TEMPO, 4-isothiocyanato-TEMPO, 4-(2-iodoacetamido)-TEMPO radical, 4-hydroxy-TEMPO, 4-cyano-TEMPO, 4-carboxy-TEMPO, 4-(2-bromoacetamido)-TEMPO, 4-amino-TEMPO or 4-acetamido-2,2,6,6-tetramethylpiperidinyl-1-oxyl. In one embodiment, the stable nitro radical compound is selected from the group consisting of TEMPO, 2,2,6,6-tetramethyl-4-(methylsulfonyloxy)-1-piperidinyloxy, 4-phosphonyloxy-TEMPO, 4-oxo-TEMPO, 4-methoxy-TEMPO, 4-isothiocyanato-TEMPO, 4-(2-iodoacetamido)-TEMPO radical, 4-hydroxy-TEMPO, 4-cyano-TEMPO, 4-carboxyl-TEMPO, 4-(2-bromoacetamido)-TEMPO, 4-amino-TEMPO, 4-acetamido-2,2,6,6-tetramethylpiperidinyl-1-oxyl. Preferably, the stable nitro radical compound is TEMPO. In another embodiment, the stable nitro radical compound is 3β-DOXYL-5α-cholestane, 5-DOXYL-stearic acid, 16-DOXYL-stearic acid, 5-DOXYL-stearate methyl ester, 3-(aminomethyl)-PROXYL, 3-aminoformyl-PROXYL, 3-aminoformyl-2,2,5,5-tetramethyl-3-pyrroline-1-oxyl, 3-carboxy-PROXYL or 3-cyano-PROXYL. In another embodiment, the stable nitro radical compound is selected from the group consisting of: 3β-DOXYL-5α-cholestane, 5-DOXYL-stearic acid, 16-DOXYL-stearic acid, 5-DOXYL-stearate methyl ester, 3-(aminomethyl)-PROXYL, 3-aminoformyl-PROXYL, 3-aminoformyl-2,2,5,5-tetramethyl-3-pyrroline-1-oxyl, 3-carboxyl-PROXYL, 3-cyano-PROXYL. In one embodiment, the oxidizing agent is a molecule with an N-halogen moiety. Preferably, the molecule is capable of selectively oxidizing a primary alcohol in the presence of a nitro radical compound. In one embodiment, the oxidizing agent is N-chlorosuccinimide, N-bromosuccinimide, N-iodosuccinimide, dichloroisocyanuric acid, 1,3,5-trichloro-1,3,5-trioxane-2,4,6-trione, dibromoisocyanuric acid, 1,3,5-tribromo-1,3,5-trioxane-2,4,6-trione, diiodoisocyanuric acid or 1,3,5-triiodo-1,3,5-trioxane-2,4,6-trione. In one embodiment, the oxidant is selected from the group consisting of: N-chlorosuccinimide, N-bromosuccinimide, N-iodosuccinimide, dichloroisocyanuric acid, 1,3,5-trichloro-1,3,5-trioxane-2,4,6-trione, dibromoisocyanuric acid, 1,3,5-tribromo-1,3,5-trioxane-2,4,6-trione, diiodoisocyanuric acid and 1,3,5-triiodo-1,3,5-trioxane-2,4,6-trione. Preferably, the oxidant is N-chlorosuccinimide.

在一態樣中，該穩定硝醯基自由基化合物為2,2,6,6-四甲基-1-哌啶基氧基自由基(TEMPO)，且該氧化劑為N-氯代丁二醯亞胺(NCS)。In one embodiment, the stable nitroxyl radical compound is 2,2,6,6-tetramethyl-1-piperidinyloxy radical (TEMPO), and the oxidizing agent is N-chlorosuccinimide (NCS).

在一實施例中，本發明係關於一種偵測肺炎鏈球菌血清型38醣結合物中之N-乙醯基-D-岩藻糖胺(D-FucNAc)及/或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在的方法，該方法包含以下步驟：a)製備肺炎鏈球菌血清型38醣結合物，及b)偵測該醣結合物中N-乙醯基-D-岩藻糖胺(D-FucNAc)及/或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。In one embodiment, the present invention relates to a method for detecting the presence of N-acetyl-D-fucosamine (D-FucNAc) and/or N-acetyl-D-quinolfuramine (D-QuiNAc) residues in a glycoconjugate of Streptococcus pneumoniae serotype 38, the method comprising the steps of: a) preparing a glycoconjugate of Streptococcus pneumoniae serotype 38, and b) detecting the presence of N-acetyl-D-fucosamine (D-FucNAc) and/or N-acetyl-D-quinolfuramine (D-QuiNAc) residues in the glycoconjugate.

在一實施例中，藉由NMR偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及/或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由2D NMR偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及/或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and/or N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by NMR. In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and/or N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by 2D NMR.

在一實施例中，藉由異核單量子相干光譜法(HSQC)、異核多鍵相關光譜法(HMBC)、相關光譜法(COSY)及/或異核單量子相干光譜法-總相關光譜法(HSQC-TOCSY)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及/或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and/or N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by heteronuclear single quantum coherence spectroscopy (HSQC), heteronuclear multibond correlation spectroscopy (HMBC), correlation spectroscopy (COSY) and/or heteronuclear single quantum coherence spectroscopy-total correlation spectroscopy (HSQC-TOCSY).

在一較佳實施例中，藉由2D 1H- 13C HSQC NMR偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及/或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。 In a preferred embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and/or N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by 2D 1 H- 13 C HSQC NMR.

在一實施例中，藉由質譜法(MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及/或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由串聯質譜法(MS/MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及/或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由氣相層析-質譜法(GC-MS)、液相層析-質譜法(LC-MS)、毛細管電泳-質譜法(CE-MS)或離子遷移譜-質譜法(IMS/MS或IMMS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及/或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由尺寸排阻層析結合質譜法(SEC/MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及/或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and/or N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by mass spectrometry (MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and/or N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by tandem mass spectrometry (MS/MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and/or N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by gas chromatography-mass spectrometry (GC-MS), liquid chromatography-mass spectrometry (LC-MS), capillary electrophoresis-mass spectrometry (CE-MS), or ion mobility shift spectrometry-mass spectrometry (IMS/MS or IMMS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and/or N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by size exclusion chromatography combined with mass spectrometry (SEC/MS).

在一實施例中，藉由氣相層析-質譜法(GC-MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及/或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由液相層析-質譜法(LC-MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及/或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由毛細管電泳-質譜法(CE-MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及/或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由離子遷移譜-質譜法(IMS/MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及/或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由親水相互作用液相層析-質譜法(HILIC-LC/MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及/或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and/or N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by gas chromatography-mass spectrometry (GC-MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and/or N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by liquid chromatography-mass spectrometry (LC-MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and/or N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by capillary electrophoresis-mass spectrometry (CE-MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and/or N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by ion mobility shift spectrometry-mass spectrometry (IMS/MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and/or N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by hydrophilic interaction liquid chromatography-mass spectrometry (HILIC-LC/MS).

在一實施例中，本發明係關於一種偵測肺炎鏈球菌血清型38醣結合物中之N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在的方法，該方法包含以下步驟：a)製備肺炎鏈球菌血清型38醣結合物，及b)偵測該醣結合物中N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。In one embodiment, the present invention relates to a method for detecting the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfuramine (D-QuiNAc) residues in a glycoconjugate of Streptococcus pneumoniae serotype 38, the method comprising the steps of: a) preparing a glycoconjugate of Streptococcus pneumoniae serotype 38, and b) detecting the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfuramine (D-QuiNAc) residues in the glycoconjugate.

在一實施例中，藉由NMR偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由2D NMR偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by NMR. In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by 2D NMR.

在一實施例中，藉由異核單量子相干光譜法(HSQC)、異核多鍵相關光譜法(HMBC)、相關光譜法(COSY)及/或異核單量子相干光譜法-總相關光譜法(HSQC-TOCSY)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by heteronuclear single quantum coherence spectroscopy (HSQC), heteronuclear multibond correlation spectroscopy (HMBC), correlation spectroscopy (COSY) and/or heteronuclear single quantum coherence spectroscopy-total correlation spectroscopy (HSQC-TOCSY).

在一較佳實施例中，藉由2D 1H- 13C HSQC NMR偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。 In a preferred embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by 2D 1 H- 13 C HSQC NMR.

在一實施例中，藉由質譜法(MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由串聯質譜法(MS/MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由氣相層析-質譜法(GC-MS)、液相層析-質譜法(LC-MS)、毛細管電泳-質譜法(CE-MS)或離子遷移譜-質譜法(IMS/MS或IMMS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由尺寸排阻層析結合質譜法(SEC/MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by mass spectrometry (MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by tandem mass spectrometry (MS/MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by gas chromatography-mass spectrometry (GC-MS), liquid chromatography-mass spectrometry (LC-MS), capillary electrophoresis-mass spectrometry (CE-MS), or ion mobility shift spectrometry-mass spectrometry (IMS/MS or IMMS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by size exclusion chromatography combined with mass spectrometry (SEC/MS).

在一實施例中，藉由氣相層析-質譜法(GC-MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由液相層析-質譜法(LC-MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由毛細管電泳-質譜法(CE-MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由離子遷移譜-質譜法(IMS/MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由親水相互作用液相層析-質譜法(HILIC-LC/MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by gas chromatography-mass spectrometry (GC-MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by liquid chromatography-mass spectrometry (LC-MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by capillary electrophoresis-mass spectrometry (CE-MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by ion mobility shift spectrometry-mass spectrometry (IMS/MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by hydrophilic interaction liquid chromatography-mass spectrometry (HILIC-LC/MS).

在一實施例中，本發明係關於一種偵測肺炎鏈球菌血清型38醣結合物中之N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在的方法，該方法包含以下步驟：a)製備肺炎鏈球菌血清型38醣結合物，及b)偵測該醣結合物中N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。In one embodiment, the present invention relates to a method for detecting the presence of an N-acetyl-D-fucosamine (D-FucNAc) residue in a Streptococcus pneumoniae serotype 38 glycoconjugate, the method comprising the steps of: a) preparing a Streptococcus pneumoniae serotype 38 glycoconjugate, and b) detecting the presence of an N-acetyl-D-fucosamine (D-FucNAc) residue in the glycoconjugate.

在一實施例中，藉由NMR偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。在一實施例中，藉由2D NMR偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) residues is detected by NMR. In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) residues is detected by 2D NMR.

在一實施例中，藉由異核單量子相干光譜法(HSQC)、異核多鍵相關光譜法(HMBC)、相關光譜法(COSY)及/或異核單量子相干光譜法-總相關光譜法(HSQC-TOCSY)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) residue is detected by heteronuclear single quantum coherence spectroscopy (HSQC), heteronuclear multibond correlation spectroscopy (HMBC), correlation spectroscopy (COSY) and/or heteronuclear single quantum coherence spectroscopy-total correlation spectroscopy (HSQC-TOCSY).

在一較佳實施例中，藉由2D 1H- 13C HSQC NMR偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。 In a preferred embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) residue is detected by 2D 1 H- 13 C HSQC NMR.

在一實施例中，藉由質譜法(MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。藉由串聯質譜法(MS/MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。在一實施例中，藉由氣相層析-質譜法(GC-MS)、液相層析-質譜法(LC-MS)、毛細管電泳-質譜法(CE-MS)或離子遷移譜-質譜法(IMS/MS或IMMS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。在一實施例中，藉由尺寸排阻層析結合質譜法(SEC/MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) residues is detected by mass spectrometry (MS). The presence of N-acetyl-D-fucosamine (D-FucNAc) residues is detected by tandem mass spectrometry (MS/MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) residues is detected by gas chromatography-mass spectrometry (GC-MS), liquid chromatography-mass spectrometry (LC-MS), capillary electrophoresis-mass spectrometry (CE-MS), or ion mobility shift spectrometry-mass spectrometry (IMS/MS or IMMS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) residues is detected by size exclusion chromatography coupled to mass spectrometry (SEC/MS).

在一實施例中，藉由氣相層析-質譜法(GC-MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。在一實施例中，藉由液相層析-質譜法(LC-MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。在一實施例中，藉由毛細管電泳-質譜法(CE-MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。在一實施例中，藉由離子遷移譜-質譜法(IMS/MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。在一實施例中，藉由親水相互作用液相層析-質譜法(HILIC-LC/MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) residues is detected by gas chromatography-mass spectrometry (GC-MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) residues is detected by liquid chromatography-mass spectrometry (LC-MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) residues is detected by capillary electrophoresis-mass spectrometry (CE-MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) residues is detected by ion mobility shift spectrometry-mass spectrometry (IMS/MS). In one embodiment, the presence of N-acetyl-D-fucosamine (D-FucNAc) residues is detected by hydrophilic interaction liquid chromatography-mass spectrometry (HILIC-LC/MS).

在一實施例中，本發明係關於一種偵測肺炎鏈球菌血清型38醣結合物中之N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在的方法，該方法包含以下步驟：a)製備肺炎鏈球菌血清型38醣結合物，及b)偵測該醣結合物中N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。In one embodiment, the present invention relates to a method for detecting the presence of an N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue in a glycoconjugate of Streptococcus pneumoniae serotype 38, the method comprising the steps of: a) preparing a glycoconjugate of Streptococcus pneumoniae serotype 38, and b) detecting the presence of an N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue in the glycoconjugate.

在一實施例中，藉由NMR偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由2D NMR偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。In one embodiment, the presence of the N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue is detected by NMR. In one embodiment, the presence of the N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue is detected by 2D NMR.

在一實施例中，藉由異核單量子相干光譜法(HSQC)、異核多鍵相關光譜法(HMBC)、相關光譜法(COSY)及/或異核單量子相干光譜法-總相關光譜法(HSQC-TOCSY)偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。In one embodiment, the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue is detected by heteronuclear single quantum coherence spectroscopy (HSQC), heteronuclear multibond correlation spectroscopy (HMBC), correlation spectroscopy (COSY) and/or heteronuclear single quantum coherence spectroscopy-total correlation spectroscopy (HSQC-TOCSY).

在一較佳實施例中，藉由2D 1H- 13C HSQC NMR偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。 In a preferred embodiment, the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue is detected by 2D 1 H- 13 C HSQC NMR.

在一實施例中，藉由質譜法(MS)偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由串聯質譜法(MS/MS)偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由氣相層析-質譜法(GC-MS)、液相層析-質譜法(LC-MS)、毛細管電泳-質譜法(CE-MS)或離子遷移譜-質譜法(IMS/MS或IMMS)偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由尺寸排阻層析結合質譜法(SEC/MS)偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。In one embodiment, the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue is detected by mass spectrometry (MS). In one embodiment, the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue is detected by tandem mass spectrometry (MS/MS). In one embodiment, the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue is detected by gas chromatography-mass spectrometry (GC-MS), liquid chromatography-mass spectrometry (LC-MS), capillary electrophoresis-mass spectrometry (CE-MS) or ion mobility shift spectrometry-mass spectrometry (IMS/MS or IMMS). In one embodiment, the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue is detected by size exclusion chromatography coupled to mass spectrometry (SEC/MS).

在一實施例中，藉由氣相層析-質譜法(GC-MS)偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由液相層析-質譜法(LC-MS)偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由毛細管電泳-質譜法(CE-MS)偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由離子遷移譜-質譜法(IMS/MS)偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。在一實施例中，藉由親水相互作用液相層析-質譜法(HILIC-LC/MS)偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。In one embodiment, the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by gas chromatography-mass spectrometry (GC-MS). In one embodiment, the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by liquid chromatography-mass spectrometry (LC-MS). In one embodiment, the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by capillary electrophoresis-mass spectrometry (CE-MS). In one embodiment, the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by ion mobility shift spectrometry-mass spectrometry (IMS/MS). In one embodiment, the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by hydrophilic interaction liquid chromatography-mass spectrometry (HILIC-LC/MS).

10 本發明之特定實施例闡述於以下編號段落 1 至 330 中 ：1. 一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目，其中Y表示絲胺酸或甘胺酸殘基且其中所有該等重複單元包含相同的Y殘基。 2. 一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目。 3. 一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目。 4. 一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目，其中Y表示絲胺酸或甘胺酸殘基，其中所有該等重複單元包含相同的Y殘基，且其中在β-D-Gal p4OAc,6(Y)之位置4處的O-乙醯基存在於約0%至約100%的該等重複單元中。 5. 如段落4之經分離之肺炎鏈球菌血清型38醣，其中該O-乙醯基存在於約50%至約100%的該等重複單元中。 6. 如段落4之經分離之肺炎鏈球菌血清型38醣，其中該O-乙醯基存在於約80%至約100%的該等重複單元中。 7. 如段落4之經分離之肺炎鏈球菌血清型38醣，其中該O-乙醯基存在於約90%至約100%的該等重複單元中。 8. 一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目，其中Y表示絲胺酸或甘胺酸殘基，其中所有該等重複單元包含相同的Y殘基，且其中在β-D-Gal p4OAc,6(Y)之位置4處的O-乙醯基存在於約100%的該等重複單元中。 9. 一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目，其中Y表示絲胺酸或甘胺酸殘基，其中所有該等重複單元包含相同的Y殘基，且其中在β-D-Gal p4OAc,6(Y)之位置4處的O-乙醯基存在於約95%的該等重複單元中。 10. 一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目且其中在β-D-Gal p4OAc,6Ser之位置4處的O-乙醯基存在於約0%至約100%的該等重複單元中。 11. 如段落10之經分離之肺炎鏈球菌血清型38醣，其中該O-乙醯基存在於約50%至約100%的該等重複單元中。 12. 如段落10之經分離之肺炎鏈球菌血清型38醣，其中該O-乙醯基存在於約80%至約100%的該等重複單元中。 13. 如段落10之經分離之肺炎鏈球菌血清型38醣，其中該O-乙醯基存在於約90%至約100%的該等重複單元中。 14. 一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目且其中在β-D-Gal p4OAc,6Ser之位置4處的O-乙醯基存在於約100%的該等重複單元中。 15. 一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目且其中在β-D-Gal p4OAc,6Ser之位置4處的O-乙醯基存在於約95%的該等重複單元中。 16. 一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目且其中在β-D-Gal p4OAc,6Gly之位置4處的O-乙醯基存在於約0%至約100%的該等重複單元中。 17. 如段落16之經分離之肺炎鏈球菌血清型38醣，其中該O-乙醯基存在於約50%至約100%的該等重複單元中。 18. 如段落16之經分離之肺炎鏈球菌血清型38醣，其中該O-乙醯基存在於約80%至約100%的該等重複單元中。 19. 如段落16之經分離之肺炎鏈球菌血清型38醣，其中該O-乙醯基存在於約90%至約100%的該等重複單元中。 20. 一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目且其中在β-D-Gal p4OAc,6Gly之位置4處的O-乙醯基存在於約100%的該等重複單元中。 21. 一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目且其中在β-D-Gal p4OAc,6Gly之位置4處的O-乙醯基存在於95%的該等重複單元中。 22. 一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目，其中Y表示絲胺酸或甘胺酸殘基且其中所有該等重複單元包含相同的Y殘基。 23. 一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目。 24. 一種具有以下重複單元之經分離之肺炎鏈球菌血清型38醣： 其中n表示重複單元之數目。 24. 如段落1至23中任一項之經分離之肺炎鏈球菌血清型38醣，其具有10與5,000個之間的重複單元。 25. 如段落1至23中任一項之經分離之肺炎鏈球菌血清型38醣，其具有50與4,500個之間的重複單元。 26. 如段落1至23中任一項之經分離之肺炎鏈球菌血清型38醣，其具有100與4,500個之間的重複單元。 27. 如段落1至23中任一項之經分離之肺炎鏈球菌血清型38醣，其具有150與2,000個之間的重複單元。 28. 如段落1至23中任一項之經分離之肺炎鏈球菌血清型38醣，其重量平均分子量在5 kDa與5000 kDa之間。 29. 如段落1至23中任一項之經分離之肺炎鏈球菌血清型38醣，其重量平均分子量在5 kDa與2000 kDa之間。 30. 如段落1至23中任一項之經分離之肺炎鏈球菌血清型38醣，其重量平均分子量在50 kDa與5000 kDa之間。 31. 如段落1至23中任一項之經分離之肺炎鏈球菌血清型38醣，其重量平均分子量在50 kDa與1000 kDa之間。 32. 如段落1至23中任一項之經分離之肺炎鏈球菌血清型38醣，其重量平均分子量在100 kDa與5000 kDa之間。 33. 如段落1至23中任一項之經分離之肺炎鏈球菌血清型38醣，其重量平均分子量在100 kDa與1000 kDa之間。 34. 如段落1至23中任一項之經分離之肺炎鏈球菌血清型38醣，其重量平均分子量在100 kDa與500 kDa之間。 35. 如段落1至23中任一項之經分離之肺炎鏈球菌血清型38醣，其重量平均分子量在300 kDa與5000 kDa之間。 36. 如段落1至23中任一項之經分離之肺炎鏈球菌血清型38醣，其重量平均分子量在300 kDa與1000 kDa之間。 37. 如段落1至23中任一項之經分離之肺炎鏈球菌血清型38醣，其重量平均分子量在500 kDa與3000 kDa之間。 38. 如段落1至23中任一項之經分離之肺炎鏈球菌血清型38醣，其重量平均分子量在500 kDa與2000 kDa之間。 39. 如段落1至23中任一項之經分離之肺炎鏈球菌血清型38醣，其重量平均分子量在500 kDa與1000 kDa之間。 40. 如段落1至23中任一項之經分離之肺炎鏈球菌血清型38醣，其大小設定為重量平均分子量在10 kDa與1000 kDa之間。 41. 如段落1至23中任一項之經分離之肺炎鏈球菌血清型38醣，其大小設定為重量平均分子量在50 kDa與500 kDa之間。 42. 如段落1至23中任一項之經分離之肺炎鏈球菌血清型38醣，其大小設定為重量平均分子量在100 kDa與400 kDa之間。 43. 一種醣結合物，其包含與載體蛋白結合之如段落1至42中任一項之經分離之肺炎鏈球菌血清型38醣。 44. 一種醣結合物，其由與載體蛋白結合之如段落1至42中任一項之經分離之肺炎鏈球菌血清型38醣組成。 45. 一種包含血清型38莢膜醣之血清型38醣結合物，其在該醣的每100個醣重複單元中包含約1至約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。 46. 如段落45之血清型38醣結合物，其在該醣的每100個醣重複單元中包含約1至約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。 46. 如段落45之血清型38醣結合物，其在該醣的每100個醣重複單元中包含約1至約65個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基。 47. 一種包含血清型38莢膜醣之血清型38醣結合物，其在該醣的每100個醣重複單元中包含約0.5至約35個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。 48. 如段落47之血清型38醣結合物，其在該醣的每100個醣重複單元中包含約0.5至約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。 49. 如段落47之血清型38醣結合物，其在該醣的每100個醣重複單元中包含約0.5至約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。 50. 一種包含血清型38莢膜醣之血清型38醣結合物，其在該醣的每100個醣重複單元中包含約1至約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約0.5至約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。 51. 一種包含血清型38莢膜醣之血清型38醣結合物，其在該醣的每100個醣重複單元中包含約1至約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約0.5至約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。 52. 一種包含血清型38莢膜醣之血清型38醣結合物，其在該醣的每100個醣重複單元中包含約1至約65個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約0.5至約32.5個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。 53. 一種包含血清型38莢膜醣之血清型38醣結合物，其在該醣的每100個醣重複單元中包含約1至約60個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約0.5至約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。 54. 一種包含血清型38莢膜醣之血清型38醣結合物，其在該醣的每100個醣重複單元中包含約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。 55. 如段落45至54中任一項之血清型38醣結合物，其包含血清型38莢膜醣，該莢膜醣包含N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基，其中D-FucNAc殘基之數目約為D-QuiNAc殘基之數目的兩倍。 56. 一種血清型38醣結合物，其包含血清型38莢膜醣，該莢膜醣包含N-乙醯基-D-岩藻糖胺(D-FucNAc)、N-乙醯基-D-奎諾糠胺(D-QuiNAc)及D-2-乙醯胺基-2,6-二去氧基-木糖-己-4-酮糖(D-Sug)殘基。 57. 如段落56之血清型38醣結合物，其包含血清型38莢膜醣，該莢膜醣在該重複單元之相同位置處包含N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基、N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基或D-2-乙醯胺基-2,6-二去氧基-木糖-己-4-酮糖(D-Sug)殘基。 58. 如段落56或57之血清型38醣結合物，其中D-FucNAc殘基之數目約為D-QuiNAc殘基之數目的兩倍。 59. 一種血清型38醣結合物，其包含具有以下重複單元之血清型38莢膜醣： 其中n表示重複單元之數目，其中Y表示絲胺酸或甘胺酸殘基，其中所有該等重複單元包含相同的Y殘基，且其中X表示N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。 60. 如段落59之血清型38醣結合物，其中該血清型38莢膜醣在該醣的每100個醣重複單元中包含約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。 61. 如段落59之血清型38醣結合物，其中該血清型38莢膜醣在該醣的每100個醣重複單元中包含約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。 62. 一種血清型38醣結合物，其包含具有以下重複單元之血清型38莢膜醣： 其中n表示重複單元之數目，其中Y表示絲胺酸或甘胺酸殘基，其中所有該等重複單元包含相同的Y殘基，且其中X表示N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。 63. 如段落62之血清型38醣結合物，其中該血清型38莢膜醣在該醣的每100個醣重複單元中包含約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。 64. 如段落62之血清型38醣結合物，其中該血清型38莢膜醣在該醣的每100個醣重複單元中包含約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。 65. 一種血清型38醣結合物，其包含具有以下重複單元之血清型38莢膜醣： 其中n表示重複單元之數目且其中X表示N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基、N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基或D-2-乙醯胺基-2,6-二去氧基-木糖-己-4-酮糖(D-Sug)殘基。 66. 如段落65之血清型38醣結合物，其中該血清型38莢膜醣在該醣的每100個醣重複單元中包含約1至約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基、約0.5至約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基及約0至約98.5個D-2-乙醯胺基-2,6-二去氧基-木糖-己-4-酮糖(D-Sug)殘基。 67. 一種血清型38醣結合物，其包含具有以下重複單元之血清型38莢膜醣： 其中n表示重複單元之數目且其中X表示N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。 68. 如段落67之血清型38醣結合物，其中該血清型38莢膜醣在該醣的每100個醣重複單元中包含約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。 69. 如段落67之血清型38醣結合物，其中該血清型38莢膜醣在該醣的每100個醣重複單元中包含約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。 70. 一種血清型38醣結合物，其包含具有以下重複單元之血清型38莢膜醣： 其中n表示重複單元之數目且其中X表示N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基、N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基或D-2-乙醯胺基-2,6-二去氧基-木糖-己-4-酮糖(D-Sug)殘基。 71. 如段落70之血清型38醣結合物，其中該血清型38莢膜醣在該醣的每100個醣重複單元中包含約1至約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基、約0.5至約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基及約0至約98.5個D-2-乙醯胺基-2,6-二去氧基-木糖-己-4-酮糖(D-Sug)殘基。 72. 一種血清型38醣結合物，其包含具有以下重複單元之血清型38莢膜醣： 其中n表示重複單元之數目且其中X表示N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。 73. 如段落72之血清型38醣結合物，其中該血清型38莢膜醣在該醣的每100個醣重複單元中包含約70個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約30個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。 74. 如段落72之血清型38醣結合物，其中該血清型38莢膜醣在該醣的每100個醣重複單元中包含約68個N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基及約32個N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基。 75. 如段落43或44之血清型38醣結合物，其中該血清型38醣結合物包含透過胺基甲酸(2-((2-側氧基乙基)硫基)乙基)酯(eTEC)間隔子與載體蛋白共價結合之血清型38醣。 75. 如段落43或44之血清型38醣結合物其中該血清型38醣結合物係藉由點擊化學製備。 76. 一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(IV)： ， 其中X係選自由CH 2(CH 2) n '、(CH 2CH 2O) mCH 2CH 2、NHCO(CH 2) n '、NHCO(CH 2CH 2O) mCH 2CH 2、OCH 2(CH 2) n '及O(CH 2CH 2O) mCH 2CH 2組成之群；其中n'係選自1至10且m係選自1至4， 其中X'係選自由CH 2O(CH 2) n''CH 2C=O、CH 2O(CH 2CH 2O) m'(CH 2) n''CH 2C=O組成之群，其中n''係選自0至10且m'係選自0至4， 其中方括號中之結構表示該血清型38醣之重複單元，且其中n表示重複單元之數目。 77. 一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(IV)，其中X為CH 2(CH 2) n '，其中n'為2，且其中X'為CH 2O(CH 2) n ''CH 2C=O，其中n''為1。 78. 一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(V)， 其中方括號中之結構表示該血清型38醣之重複單元，且其中n表示重複單元之數目。 79. 一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(XII)： ， 其中X係選自由CH 2(CH 2) n '、(CH 2CH 2O) mCH 2CH 2、NHCO(CH 2) n '、NHCO(CH 2CH 2O) mCH 2CH 2、OCH 2(CH 2) n '及O(CH 2CH 2O) mCH 2CH 2組成之群；其中n'係選自0至10且m係選自1至4， 其中X'係選自由CH 2(CH 2) n ''、CH 2O(CH 2) n ''CH 2、CH 2O(CH 2CH 2O) m '(CH 2) n ''CH 2組成之群，其中n''係選自0至10且m'係選自0至4，且其中方括號中之結構表示該血清型38醣之重複單元，且其中n表示重複單元之數目。 80. 一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(IV)，其中X為CH 2(CH 2) n '，其中n'為0，且其中X'為CH 2(CH 2) n ''，其中n''為0。 81. 一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(XIII)， 其中方括號中之結構表示該血清型38醣之重複單元，且其中n表示重複單元之數目。 80. 如段落45至81中任一項之醣結合物，其中該血清型38醣為如段落1至42中任一項之經分離之肺炎鏈球菌血清型38醣。 81. 如段落43至81中任一項之醣結合物，其中在結合之前該肺炎鏈球菌血清型38醣之重量平均分子量(Mw)在50 kDa與1,000 kDa之間。 82. 如段落43至81中任一項之醣結合物，其中在結合之前該肺炎鏈球菌血清型38醣之重量平均分子量(Mw)在100 kDa與600 kDa之間。 83. 如段落43至81中任一項之醣結合物，其中在結合之前該肺炎鏈球菌血清型38醣之重量平均分子量(Mw)在100 kDa與400 kDa之間。 84. 如段落43至81中任一項之醣結合物，其中在結合之前該肺炎鏈球菌血清型38醣之重量平均分子量(Mw)在150 kDa與300 kDa之間。 85. 如段落43至84中任一項之醣結合物，其中該血清型38醣結合物之重量平均分子量(Mw)在250 kDa與20,000 kDa之間。 86. 如段落43至84中任一項之醣結合物，其中該血清型38醣結合物之重量平均分子量(Mw)在500 kDa與15,000 kDa之間。 87. 如段落43至84中任一項之醣結合物，其中該血清型38醣結合物之重量平均分子量(Mw)在500 kDa與10,000 kDa之間。 87a. 如段落43至84中任一項之醣結合物，其中該血清型38醣結合物之重量平均分子量(Mw)在1,000 kDa與10,000 kDa之間。 87b. 如段落43至81中任一項之醣結合物，其中在結合之前該肺炎鏈球菌血清型38醣之重量平均分子量(Mw)在150 kDa與300 kDa之間，且其中該血清型38醣結合物之重量平均分子量(Mw)在1,000 kDa與10,000 kDa之間。 88. 如段落43至84中任一項之醣結合物，其中該血清型38醣結合物之重量平均分子量(Mw)在750 kDa與7,500 kDa之間。 89. 如段落43至84中任一項之醣結合物，其中該血清型38醣結合物之重量平均分子量(Mw)在1,000 kDa與5,000 kDa之間。 90. 如段落43至89中任一項之醣結合物，其中該血清型38醣結合物之結合程度在2與15之間。 91. 如段落43至89中任一項之醣結合物，其中該血清型38醣結合物之結合程度在2與10之間。 92. 如段落43至89中任一項之醣結合物，其中該血清型38醣結合物之結合程度在3與5之間。 93. 如段落43至89中任一項之醣結合物，其中該血清型38醣結合物之結合程度在2與6之間。 94. 如段落43至89中任一項之醣結合物，其中該血清型38醣結合物之結合程度在4與10之間。 94a. 如段落43至81中任一項之醣結合物，其中在結合之前該肺炎鏈球菌血清型38醣之重量平均分子量(Mw)在150 kDa與300 kDa之間，其中該血清型38醣結合物之重量平均分子量(Mw)在1,000 kDa與10,000 kDa之間且其中該血清型38醣結合物之結合程度在2與10之間。 94b. 如段落43至81中任一項之醣結合物，其中在結合之前該肺炎鏈球菌血清型38醣之重量平均分子量(Mw)在150 kDa與300 kDa之間，其中該血清型38醣結合物之重量平均分子量(Mw)在1,000 kDa與10,000 kDa之間且其中該血清型38醣結合物之結合程度在4與10之間。 95. 如段落43至94中任一項之醣結合物，其中該醣結合物中之血清型38醣與載體蛋白之比率(w/w)在0.5與3.0之間。 97. 如段落43至94中任一項之醣結合物，其中該醣結合物中之血清型38醣與載體蛋白之比率(w/w)在0.5與2.0之間。 98. 如段落43至94中任一項之醣結合物，其中該醣結合物中之血清型38醣與載體蛋白之比率(w/w)在0.5與1.5之間。 99. 如段落43至94中任一項之醣結合物，其中該醣結合物中之血清型38醣與載體蛋白之比率(w/w)在0.8與1.2之間。 100. 如段落43至94中任一項之醣結合物，其中該醣結合物中之血清型38醣與載體蛋白之比率(w/w)在0.5與1.0之間。 101. 如段落43至94中任一項之醣結合物，其中該醣結合物中之血清型38醣與載體蛋白之比率(w/w)在1.0與1.5之間。 102. 如段落43至94中任一項之醣結合物，其中該醣結合物中之血清型38醣與載體蛋白之比率(w/w)在1.0與2.0之間。 103. 如段落43至94中任一項之醣結合物，其中該醣結合物中之血清型38醣與載體蛋白之比率(w/w)在0.8與1.2之間。 104. 如段落43至94中任一項之醣結合物，其中該醣結合物中之血清型38醣與載體蛋白之比率(w/w)在0.7與1.1之間。 104a. 如段落43至81中任一項之醣結合物，其中在結合之前該肺炎鏈球菌血清型38醣之重量平均分子量(Mw)在150 kDa與300 kDa之間，其中該血清型38醣結合物之重量平均分子量(Mw)在1,000 kDa與10,000 kDa之間，其中該醣結合物中之血清型38醣與載體蛋白之比率(w/w)在0.5與1.5之間且其中該血清型38醣結合物之結合程度在4與10之間。 104b. 如段落43至81中任一項之醣結合物，其中在結合之前該肺炎鏈球菌血清型38醣之重量平均分子量(Mw)在150 kDa與300 kDa之間，其中該血清型38醣結合物之重量平均分子量(Mw)在1,000 kDa與10,000 kDa之間，其中該醣結合物中之血清型38醣與載體蛋白之比率(w/w)在0.7與1.1之間且其中該血清型38醣結合物之結合程度在4與10之間。 105. 如段落43至104中任一項之醣結合物，其中該血清型38醣結合物包含與血清型38醣之總量相比小於約50%之游離血清型38醣。 106. 如段落43至104中任一項之醣結合物，其中該血清型38醣結合物包含與血清型38醣之總量相比小於約25%之游離血清型38醣。 107. 如段落43至104中任一項之醣結合物其中該血清型38醣結合物包含與血清型38醣之總量相比小於約20%之游離血清型38醣。 108. 如段落43至104中任一項之醣結合物其中該血清型38醣結合物包含與血清型38醣之總量相比小於約15%之游離血清型38醣。 108a. 如段落43至81中任一項之醣結合物，其中在結合之前該肺炎鏈球菌血清型38醣之重量平均分子量(Mw)在150 kDa與300 kDa之間，其中該血清型38醣結合物之重量平均分子量(Mw)在1,000 kDa與10,000 kDa之間，其中該醣結合物中之血清型38醣與載體蛋白之比率(w/w)在0.5與1.5之間，其中該血清型38醣結合物之結合程度在4與10之間且其中該血清型38醣結合物包含與血清型38醣之總量相比小於約20%之游離血清型38醣。 108b. 如段落43至81中任一項之醣結合物，其中在結合之前該肺炎鏈球菌血清型38醣之重量平均分子量(Mw)在150 kDa與300 kDa之間，其中該血清型38醣結合物之重量平均分子量(Mw)在1,000 kDa與10,000 kDa之間，其中該醣結合物中之血清型38醣與載體蛋白之比率(w/w)在0.7與1.1之間，其中該血清型38醣結合物之結合程度在4與10之間且其中該血清型38醣結合物包含與血清型38醣之總量相比小於約20%之游離血清型38醣。 109. 如段落43至108中任一項之醣結合物，其中至少30%的該血清型38醣結合物在CL-4B管柱中之K d低於或等於0.3。 110. 如段落43至108中任一項之醣結合物，其中至少40%的該醣結合物在CL-4B管柱中之K d低於或等於0.3。 111. 如段落43至108中任一項之醣結合物，其中至少60%的該血清型38醣結合物在CL-4B管柱中之K d低於或等於0.3。 112. 如段落43至108中任一項之醣結合物，其中50%與80%之間的該血清型38醣結合物在CL-4B管柱中之K d低於或等於0.3。 113. 如段落43至108中任一項之醣結合物，其中65%與80%之間的該血清型38醣結合物在CL-4B管柱中之K d低於或等於0.3。 113a. 如段落43至81中任一項之醣結合物，其中在結合之前該肺炎鏈球菌血清型38醣之重量平均分子量(Mw)在150 kDa與300 kDa之間，其中該血清型38醣結合物之重量平均分子量(Mw)在1,000 kDa與10,000 kDa之間，其中該醣結合物中之血清型38醣與載體蛋白之比率(w/w)在0.5與1.5之間，其中該血清型38醣結合物之結合程度在4與10之間，其中該血清型38醣結合物包含與血清型38醣之總量相比小於約20%之游離血清型38醣且其中65%與80%之間的該血清型38醣結合物在CL-4B管柱中之K d低於或等於0.3。 113b. 如段落43至81中任一項之醣結合物，其中在結合之前該肺炎鏈球菌血清型38醣之重量平均分子量(Mw)在150 kDa與300 kDa之間，其中該血清型38醣結合物之重量平均分子量(Mw)在1,000 kDa與10,000 kDa之間，其中該醣結合物中之血清型38醣與載體蛋白之比率(w/w)在0.7與1.1之間，其中該血清型38醣結合物之結合程度在4與10之間，其中該血清型38醣結合物包含與血清型38醣之總量相比小於約20%之游離血清型38醣且其中65%與80%之間的該血清型38醣結合物在CL-4B管柱中之K d低於或等於0.3。 114. 如段落43至113中任一項之醣結合物其中該血清型38醣結合物之該載體蛋白係選自由以下組成之群：TT、DT、DT突變體及來自鏈球菌之C5a肽酶(SCP)。 115. 如段落43至113中任一項之醣結合物，其中該血清型38醣結合物之該載體蛋白係根瘤菌抗生物素蛋白[aa 45-179J-GGGGSSS-SP1500- AAA-SP0785] (CP1)。 115a. 如段落43至113中任一項之醣結合物，其中該血清型38醣結合物之該載體蛋白為Rhavi-連接子-PdT(G294P)-連接子-SP0435 [aa 62-185]融合蛋白(SPP2)。 116. 如段落43至113中任一項之醣結合物，其中該血清型38醣結合物之該載體蛋白為DT。 117. 如段落43至113中任一項之醣結合物，其中該血清型38醣結合物之該載體蛋白為TT。 118. 如段落43至113中任一項之醣結合物，其中該血清型38醣結合物之該載體蛋白為PD。 119. 如段落43至113中任一項之醣結合物，其中該血清型38醣結合物之該載體蛋白為CRM 197或來自鏈球菌之C5a肽酶(SCP)。 120. 如段落43至113中任一項之醣結合物，其中該血清型38醣結合物之該載體蛋白為CRM 197。 121. 如段落43至113中任一項之醣結合物，其中該血清型38醣結合物之該載體蛋白為SCP。 122. 如段落43至113中任一項之醣結合物，其中該血清型38醣結合物之該載體蛋白為酶不活化的SCP。 123. 如段落43至113中任一項之醣結合物，其中該血清型38醣結合物之該載體蛋白為酶不活化的來自GBS之SCP (SCPB)。 124. 如段落43至113中任一項之醣結合物，其中該血清型38醣結合物之該載體蛋白為酶不活化的來自GAS之SCP (SCPA)。 125. 如段落43至113中任一項之醣結合物，其中該血清型38醣結合物之該之載體蛋白為SCPB之片段。 126. 如段落43至113中任一項之醣結合物，其中該血清型38醣結合物之該載體蛋白係由SEQ ID NO: 1組成的酶不活化的SCP之片段。 127. 如段落43至113中任一項之醣結合物，其中該血清型38醣結合物之該載體蛋白係由SEQ ID NO: 2組成的酶不活化的SCP之片段。 128. 一種免疫原組合物，其包含如段落1至42中任一項之肺炎鏈球菌血清型38醣。 129. 一種免疫原組合物，其包含如段落43至127中任一項之肺炎鏈球菌血清型38醣結合物。 130. 如段落129之免疫原組合物，其包含1至45種不同醣結合物。 131. 如段落129之免疫原組合物，其包含來自1至45種不同肺炎鏈球菌血清型之醣結合物。 132. 如段落129之免疫原組合物，其包含來自7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24或25種不同肺炎鏈球菌血清型之醣結合物。 133. 如段落129之免疫原組合物，其包含來自16或20種不同肺炎鏈球菌血清型之醣結合物。 134. 如段落129之免疫原組合物，其為21、22、23、24或25價肺炎鏈球菌結合物組合物。 135. 如段落129之免疫原組合物，其為21價肺炎鏈球菌結合物組合物。 136. 如段落129之免疫原組合物，其為22價肺炎鏈球菌結合物組合物。 137. 如段落129之免疫原組合物，其為23價肺炎鏈球菌結合物組合物。 138. 如段落129之免疫原組合物，其為24價肺炎鏈球菌結合物組合物。 139. 如段落129之免疫原組合物，其為25價肺炎鏈球菌結合物組合物。 140. 如段落129之免疫原組合物，其包含來自26至45種不同肺炎鏈球菌血清型之醣結合物的醣結合物。 141. 如段落129之免疫原組合物，其包含來自26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、41、42、43、44或45種不同肺炎鏈球菌血清型之醣結合物。 142. 如段落129之免疫原組合物，其包含來自35或45種不同肺炎鏈球菌血清型之醣結合物。 142a. 如段落129之免疫原組合物，其包含來自35種不同肺炎鏈球菌血清型之醣結合物。 143. 如段落129之免疫原組合物，其為35、36、37、38、39、40、41、42、43、44或45價肺炎鏈球菌結合物組合物。 143a. 如段落129之免疫原組合物，其為35價肺炎鏈球菌結合物組合物。 144. 如段落129之免疫原組合物，其為40、41、42、43、44或45價肺炎鏈球菌結合物組合物。 145. 如段落129之免疫原組合物，其為40價肺炎鏈球菌結合物組合物。 146. 如段落129之免疫原組合物，其為41價肺炎鏈球菌結合物組合物。 147. 如段落129之免疫原組合物，其為42價肺炎鏈球菌結合物組合物。 148. 如段落129之免疫原組合物，其為43價肺炎鏈球菌結合物組合物。 149. 如段落129之免疫原組合物，其為44價肺炎鏈球菌結合物組合物。 150. 如段落129之免疫原組合物，其為45價肺炎鏈球菌結合物組合物。 151. 如段落129至150中任一項之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型4、6B、9V、14、18C、19F及23F之醣結合物。 152. 如段落151之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型1、5及7F之醣結合物。 153. 如段落152之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型3之醣結合物。 154. 如段落153之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型6A及19A之醣結合物。 155. 如段落154之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型22F及33F之醣結合物。 156. 如段落155之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型8、10A、11A、12F及15B之醣結合物。 157. 如段落156之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型2之醣結合物。 158. 如段落157之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型9N之醣結合物。 159. 如段落158之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型17F之醣結合物。 160. 如段落159之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型20之醣結合物。 161. 如段落160之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型2之醣結合物。 162. 如段落161之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型15C之醣結合物。 163.如段落129至150中任一項之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型4、6B、9V、14、18C、19F及23F之醣結合物，且其中該免疫原組合物為8價肺炎鏈球菌結合物組合物。 164. 如段落129至150中任一項之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型1、4、5、6B、7F、9V、14、18C、19F及23F之醣結合物，且其中該免疫原組合物為11價肺炎鏈球菌結合物組合物。 165. 如段落129至150中任一項之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型1、3、4、5、6A、6B、7F、9V、14、18C、19A、19F及23F之醣結合物，且其中該免疫原組合物為14價肺炎鏈球菌結合物組合物。 166. 如段落129至150中任一項之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型1、3、4、5、6A、6B、7F、9V、14、18C、19A、19F、22F、23F及33F之醣結合物，且其中該免疫原組合物為16價肺炎鏈球菌結合物組合物。 167. 如段落129至150中任一項之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型1、3、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15B、18C、19A、19F、22F、23F之醣結合物，且其中該免疫原組合物為21價肺炎鏈球菌結合物組合物。 168. 如段落129至150中任一項之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型1、3、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15C、18C、19A、19F、22F、23F及33F之醣結合物。 169. 如段落168之免疫原組合物，其為21價肺炎鏈球菌結合物組合物。 170. 如段落129至150中任一項之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型1、2、3、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15B、18C、19A、19F、22F、23F及33F之醣結合物。 171. 如段落170之免疫原組合物，其為22價肺炎鏈球菌結合物組合物。 172. 如段落129至150中任一項之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型1、2、3、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15C、18C、19A、19F、22F、23F及33F之醣結合物。 173. 如段落172之免疫原組合物，其為22價肺炎鏈球菌結合物組合物。 174. 如段落129至150中任一項之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型1、3、4、5、6A、6B、7F、8、9V、9N、10A、11A、12F、14、15B、18C、19A、19F、22F、23F及33F之醣結合物。 175. 如段落174之免疫原組合物，其為22價肺炎鏈球菌結合物組合物。 176. 如段落129至150中任一項之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型1、2、3、4、5、6A、6B、7F、8、9V、9N、10A、11A、12F、14、15B、18C、19A、19F、22F、23F及33F之醣結合物。 177. 如段落176之免疫原組合物，其為23價肺炎鏈球菌結合物組合物。 178. 如段落129至150中任一項之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型1、2、3、4、5、6A、6B、7F、8、9V、9N、10A、11A、12F、14、15B、17F、18C、19A、19F、22F、23F及33F之醣結合物。 179. 如段落178之免疫原組合物，其為24價肺炎鏈球菌結合物組合物。 180. 如段落129至150中任一項之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型1、2、3、4、5、6A、6B、7F、8、9V、9N、10A、11A、12F、14、15B、17F、18C、19A、19F、20、22F、23F及33F之醣結合物。 181. 如段落180之免疫原組合物，其為25價肺炎鏈球菌結合物組合物。 182. 如段落129至150中任一項之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型1、3、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15A、15B、18C、19A、19F、22F、23F及33F之醣結合物。 183. 如段落182之免疫原組合物，其為22價肺炎鏈球菌結合物組合物。 184. 如段落129至150中任一項之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型1、3、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15B、18C、19A、19F、22F、23A、23F及33F之醣結合物。 185. 如段落184之免疫原組合物，其為22價肺炎鏈球菌結合物組合物。 186. 如段落129至150中任一項之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型1、3、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15B、18C、19A、19F、22F、23B、23F及33F之醣結合物。 187. 如段落186之免疫原組合物，其為22價肺炎鏈球菌結合物組合物。 188. 如段落129至150中任一項之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型1、3、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15B、18C、19A、19F、22F、23F、24F及33F之醣結合物。 189. 如段落188之免疫原組合物，其為22價肺炎鏈球菌結合物組合物。 190. 如段落129至150中任一項之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型1、3、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15B、18C、19A、19F、22F、23F、33F及35B之醣結合物。 191. 如段落190之免疫原組合物，其為22價肺炎鏈球菌結合物組合物。 192. 如段落129至150中任一項之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型1、3、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15A、15B、18C、19A、19F、22F、23A、23F及33F之醣結合物。 193. 如段落192之免疫原組合物，其為23價肺炎鏈球菌結合物組合物。 194. 如段落129至150中任一項之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型1、3、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15A、15B、18C、19A、19F、22F、23A、23B、23F、24F、33F及35B之醣結合物。 195. 如段落194之免疫原組合物，其中該肺炎鏈球菌醣與CRM 197結合。 196. 如段落194之免疫原組合物，其中來自血清型1、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15A、15B、18C、19A、19F、22F、23A、23B、23F、24F、33F、35B及38之該肺炎鏈球菌醣與CRM 197結合且該肺炎鏈球菌醣血清型3與SCP結合。 197. 如段落196之免疫原組合物，其為26價肺炎鏈球菌結合物組合物。 198. 如段落129至150中任一項之免疫原組合物，其進一步包含至少一種選自由以下組成之群之醣結合物：來自肺炎鏈球菌血清型2、3、7C、9N、10B、15A、16F、17F、20、21、22A、23A、23B、24B、24F、27、29、31、33B、34、35B及35F之醣結合物。 199. 如段落129至150中任一項之免疫原組合物，其進一步包含二十一種選自由以下組成之群之醣結合物：來自肺炎鏈球菌血清型2、3、7C、9N、10B、15A、16F、17F、20、21、22A、23A、23B、24B、24F、27、29、31、33B、34、35B及35F之醣結合物。 200. 如段落199之免疫原組合物，其為22價肺炎鏈球菌結合物組合物。 201. 如段落129至150中任一項之免疫原組合物，其進一步包含來自血清型2、3、7C、9N、10B、15A、16F、17F、20、21、22A、23A、23B、24B、24F、27、29、31、33B、34、35B及35之醣結合物。 202. 如段落201之免疫原組合物，其為23價肺炎鏈球菌結合物組合物。 203. 如段落129至150中任一項之免疫原組合物，其進一步包含至少一種選自由以下組成之群之醣結合物：來自肺炎鏈球菌血清型2、3、7C、9N、10B、15A、16F、17F、19A、19F、20、21、22A、23A、23B、24B、24F、27、29、31、33B、34、35B及35F之醣結合物。 204. 如段落129至150中任一項之免疫原組合物，其進一步包含二十二種選自由以下組成之群之醣結合物：來自肺炎鏈球菌血清型2、3、7C、9N、10B、15A、16F、17F、19A、19F、20、21、22A、23A、23B、24B、24F、27、29、31、33B、34、35B及35F之醣結合物。 205. 如段落204之免疫原組合物，其為23價肺炎鏈球菌結合物組合物。 206. 如段落129至150中任一項之免疫原組合物，其進一步包含二十三種選自由以下組成之群之醣結合物：來自肺炎鏈球菌血清型2、3、7C、9N、10B、15A、16F、17F、19A、19F、20、21、22A、23A、23B、24B、24F、27、29、31、33B、34、35B及35F之醣結合物。 207. 如段落206之免疫原組合物，其為24價肺炎鏈球菌結合物組合物。 208. 如段落129至150中任一項之免疫原組合物，其進一步包含來自血清型2、3、7C、9N、10B、15A、16F、17F、20、21、22A、23A、23B、24B、24F、27、29、31、33B、34、35B及35F之醣結合物。 209. 如段落208之免疫原組合物，其為23價肺炎鏈球菌結合物組合物。 210. 如段落129至150中任一項之免疫原組合物，其進一步包含來自血清型2、3、7C、9N、10B、15A、16F、17F、19A、19F、20、21、22A、23A、23B、24B、24F、27、29、31、33B、34、35B及35F之醣結合物。 211. 如段落210之免疫原組合物，其為23價肺炎鏈球菌結合物組合物。 212. 如段落128至211中任一項之免疫原組合物，其進一步包含至少一種、兩種或三種佐劑。 213. 如段落128至211中任一項之免疫原組合物，其進一步包含一種佐劑。 214. 如段落128至211中任一項之免疫原組合物，其進一步包含兩種佐劑。 215. 如段落128至211中任一項之免疫原組合物，其進一步包含作為佐劑之鋁鹽(明礬)。 216. 如段落128至211中任一項之免疫原組合物，其進一步包含作為佐劑之磷酸鋁。 217. 如段落128至211中任一項之免疫原組合物，其進一步包含基於皂素之佐劑。 218. 如段落128至211中任一項之免疫原組合物，進一步包含基於QS21之佐劑。 219. 如段落128至211中任一項之免疫原組合物，其進一步包含作為佐劑之CpG寡核苷酸。 220. 如段落128至219中任一項之免疫原組合物，其用作疫苗。 221. 如段落128至219中任一項之免疫原組合物，其用於預防、治療或改善個體之與肺炎鏈球菌血清型38相關之感染、疾病或病況的方法中。 222. 如段落128至219中任一項之免疫原組合物，其用於誘導個體中之針對肺炎鏈球菌血清型38之免疫反應的方法中，該方法包含向該個體投與免疫有效量之該免疫原組合物。 223. 如段落128至219中任一項之免疫原組合物，其用於預防個體中受肺炎鏈球菌38感染之方法中，該方法包含向該個體投與免疫有效量之該免疫原組合物。 224. 一種偵測經分離之肺炎鏈球菌血清型38多醣中之D-2-乙醯胺基-2,6-二去氧基-木糖-己-4-酮糖(D-Sug)殘基之存在的方法，該方法包含以下步驟：a)分離肺炎鏈球菌血清型38多醣及b)偵測該多醣中D-2-乙醯胺基-2,6-二去氧基-木糖-己-4-酮糖殘基之存在。 225. 如段落224之方法，其中藉由NMR或質譜法(MS)偵測到D-Sug殘基之存在。 226. 一種偵測經還原之血清型38多醣中之N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在的方法，該方法包含以下步驟：a)使經分離之肺炎鏈球菌血清型38多醣與還原劑反應，及b)偵測該經還原之多醣中N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。 226. 如段落225之方法，其中藉由NMR偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。 227. 如225至226中任一項之方法，其中該還原劑為硼氫化鈉(NaBH 4)。 228. 如225至227中任一項之方法，其中該經分離之肺炎鏈球菌血清型38多醣先前已用氧化劑處理。 229. 一種偵測經還原之血清型38多醣中之N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在的方法，該方法包含以下步驟：a)使經分離之肺炎鏈球菌血清型38多醣與還原劑反應，及b)偵測該經還原之多醣中N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。 230. 如段落229之方法，其中藉由NMR偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。 231. 如段落229之方法，其中藉由質譜法(MS)偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。 232. 如229至231中任一項之方法，其中該還原劑為硼氫化鈉(NaBH 4)。 233. 如229至232中任一項之方法，其中該經分離之肺炎鏈球菌血清型38多醣先前已用氧化劑處理。 234. 一種偵測經還原之血清型38多醣中之N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在的方法，該方法包含以下步驟：a)使經分離之肺炎鏈球菌血清型38多醣與還原劑反應，及b)偵測該經還原之多醣中N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。 235. 如段落234之方法，其中藉由NMR偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。 236. 如段落234之方法，其中藉由質譜法(MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。 237. 如234至236中任一項之方法，其中該還原劑為硼氫化鈉(NaBH 4)。 238. 如234至237中任一項之方法，其中該經分離之肺炎鏈球菌血清型38多醣先前已用氧化劑處理。 239. 一種偵測肺炎鏈球菌血清型38醣結合物中之N-乙醯基-D-岩藻糖胺(D-FucNAc)及/或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在的方法，該方法包含以下步驟：a)製備肺炎鏈球菌血清型38醣結合物，及b)偵測該醣結合物中N-乙醯基-D-岩藻糖胺(D-FucNAc)及/或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。 240. 如段落239之方法，其中藉由NMR偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及/或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。 241. 如段落239之方法，其中藉由質譜法(MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及/或N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。 242. 一種偵測肺炎鏈球菌血清型38醣結合物中之N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在的方法，該方法包含以下步驟：a)製備肺炎鏈球菌血清型38醣結合物，及b)偵測該醣結合物中N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。 243. 如段落242之方法，其中藉由NMR偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。 244. 如段落242之方法，其中藉由質譜法(MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)及N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。 245. 一種偵測肺炎鏈球菌血清型38醣結合物中之N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在的方法，該方法包含以下步驟：a)製備肺炎鏈球菌血清型38醣結合物，及b)偵測該醣結合物中N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。 246. 如段落245之方法，其中藉由NMR偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。 247. 如段落245之方法，其中藉由質譜法(MS)偵測到N-乙醯基-D-岩藻糖胺(D-FucNAc)殘基之存在。 248. 一種偵測肺炎鏈球菌血清型38醣結合物中之N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在的方法，該方法包含以下步驟：a)製備肺炎鏈球菌血清型38醣結合物，及b)偵測該醣結合物中N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。 249. 如段落248之方法，其中藉由NMR偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。 250. 如段落248之方法，其中藉由質譜法(MS)偵測到N-乙醯基-D-奎諾糠胺(D-QuiNAc)殘基之存在。 251.  如段落43至127中任一項之醣結合物，其用作抗原。 252.  如段落43至127中任一項之醣結合物，其用作藥劑。 253.  如段落43至127中任一項之醣結合物，其用作疫苗。 254   如段落43至127中任一項之醣結合物，其用於在個體中產生免疫反應。 255.  如段落254所使用之醣結合物，其中該個體為人類。 256.  如段落128至219中任一項之免疫原組合物，其用作藥劑。 257.  如段落128至219中任一項之免疫原組合物，其用作疫苗。 258.  如段落257所使用之免疫原組合物，其中該個體為人類。 259.  如段落128至219中任一項之免疫原組合物，其用於預防、治療或改善個之細菌性感染、疾病或病況的方法。 260.  如段落128至219中任一項之免疫原組合物，其用於預防、治療或改善個體之肺炎鏈球菌血清型38感染、疾病或病況的方法。 261.  如段落128至219中任一項之免疫原組合物，其用於誘導針對個體之肺炎鏈球菌血清型38之免疫反應的方法。 262.  如段落128至219中任一項之免疫原組合物，其用於預防個體之肺炎鏈球菌血清型38感染的方法。 263.  如段落128至219中任一項之免疫原組合物，其用於保護易受肺炎鏈球菌血清型38感染影響之人類的方法。 264.  一種使用點擊化學製備肺炎鏈球菌血清型38醣結合物之方法，該方法包含以下步驟：(a)使經分離血清型38醣與碳酸衍生物及疊氮基連接子在非質子性溶劑中反應以產生經活化之疊氮基醣(醣的活化)，(b)使載體蛋白與帶有N-羥基丁二醯亞胺(NHS)部分和炔烴基之試劑反應，其中NHS部分與胺基反應以形成醯胺鍵，從而獲得炔烴官能化載體蛋白(載體蛋白的活化)，(c)藉由Cu+1介導之疊氮化物-炔烴環加成反應使步驟(a)之經活化之疊氮基醣與步驟(b)之經活化的炔烴-載體蛋白反應以形成醣結合物。 265.  如段落264之方法，其中在活化(a)之前，已將該醣之尺寸設定為目標分子量(MW)範圍。 266.  如段落264之方法，其中該經分離之血清型38醣在用碳酸衍生物及疊氮基連接子活化之前經尺寸設定。 267.  如段落266之方法，其中該經分離之血清型38莢膜多醣之尺寸設定為重量平均分子量在50 kDa與500 kDa之間。 268.  如段落266之方法，其中該經分離之血清型38莢膜多醣之尺寸設定為重量平均分子量在50 kDa與400 kDa之間。 268.  如段落266之方法，其中該經分離之血清型38莢膜多醣之尺寸設定為重量平均分子量在100 kDa與500 kDa之間。 270.  如段落266之方法，其中該經分離之血清型38莢膜多醣之尺寸設定為重量平均分子量在約100 kDa與約400 kDa之間。 271.  如段落266之方法，其中該經分離之血清型38莢膜多醣之尺寸設定為重量平均分子量在約150 kDa與約300 kDa之間。 272.  如段落264之方法，其中該經分離之血清型38莢膜多醣在用碳酸衍生物及疊氮基連接子活化之前未經尺寸設定。 273.  如段落264至272中任一項之方法，其中該碳酸衍生物係選自由以下組成之群：1,1'-羰基二咪唑(CDI)、1,1'-羰基-二-(1,2,4-三唑) (CDT)、碳酸N,N'-二丁二醯亞胺酯(DSC)及氯甲酸N-羥基丁二醯亞胺酯。 274.  如段落264至272中任一項之方法，其中該碳酸衍生物為1,1'-羰基二咪唑(CDI)。 275.  如段落264至272中任一項之方法，其中該碳酸衍生物為1,1'-羰基-二-(1,2,4-三唑) (CDT)。 276.  如段落264至272中任一項之方法，其中該碳酸衍生物為碳酸N,N'-二丁二醯亞胺酯(DSC)。 277.  如段落264至272中任一項之方法，其中該碳酸衍生物為氯甲酸N-羥基丁二醯亞胺酯。 278.  如段落264至277中任一項之方法，其中該疊氮基連接子為式(VI)之化合物， 其中X係選自由CH 2(CH 2) n、(CH 2CH 2O) mCH 2CH 2、NHCO(CH 2) n、NHCO(CH 2CH 2O) mCH 2CH 2、OCH 2(CH2) n及O(CH 2CH 2O) mCH 2CH 2組成之群；其中n係選自1至10且m係選自1至4。 279.  如段落264至277中任一項之方法，其中該疊氮基連接子為式(VI)之化合物，其中X為CH 2(CH 2) n，且n係選自1至10。 280.  如段落264至277中任一項之方法，其中該疊氮基連接子為式(VI)之化合物，其中X為(CH 2CH 2O) mCH 2CH 2，其中m係選自1至4。 281.  如段落264至277中任一項之方法，其中該疊氮基連接子為式(VI)之化合物，其中X為NHCO(CH 2) n，且n係選自1至10。 282.  如段落264至277中任一項之方法，其中該疊氮基連接子為式(VI)之化合物，其中X為NHCO(CH 2CH 2O) mCH 2CH 2，其中m係選自1至4。 283.  如段落264至277中任一項之方法，其中該疊氮基連接子為式(VI)之化合物，其中X為OCH 2(CH 2) n，且n係選自1至10。 284.  如段落264至277中任一項之方法，其中該疊氮基連接子為式(VI)之化合物，其中X為O(CH 2CH 2O) mCH 2CH 2，其中m係選自1至4。 285.  如段落264至277中任一項之方法，其中該疊氮基連接子為式(VII)之化合物， 。 286.  如段落264至277中任一項之方法，其中該疊氮基連接子為3-疊氮基-丙胺。 287.  如段落264至286中任一項之方法，其中該帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑為帶有N-羥基丁二醯亞胺(NHS)部分及末端炔烴之試劑。 288.  如段落264至286中任一項之方法，其中該帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑為帶有N-羥基丁二醯亞胺(NHS)部分及環炔烴之試劑。 289.  如段落264至286中任一項之方法，其中該帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑為式(VIII)之化合物， 其中X係選自由CH 2O(CH 2) nCH 2C=O及CH 2O(CH 2CH 2O) m(CH 2) nCH 2C=O組成之群，其中n係選自0至10且m係選自0至4。 290.  如段落264至286中任一項之方法，其中該帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑為式(VIII)之化合物，其中X為CH 2O(CH 2) nCH 2C=O，其中n係選自0至10。 291.  如段落264至286中任一項之方法，其中該帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑為式(VIII)之化合物，其中X為CH2O(CH2CH2O)m(CH2)nCH2C=O，其中n係選自0至10且m係選自0至4。 292.  如段落264至286中任一項之方法，其中該帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑為式(IX)之化合物： 。 293.  如段落264至292中任一項之方法，其中步驟a)包含使醣與碳酸衍生物反應，隨後使經碳酸衍生物活化之醣與疊氮基連接子在非質子性溶劑中反應以產生經活化之疊氮基醣。 294.  如段落264至293中任一項之方法，其中步驟a)包含使醣與相對於反應混合物中所存在之醣的量在0.01至10莫耳當量之間的量的碳酸衍生物反應。 295.  如段落264至294中任一項之方法，其中在步驟a)處，使經分離醣與碳酸衍生物在非質子性溶劑中反應。 296.  如段落264至295中任一項之方法，其中使經分離醣與CDI在包含0.1%至1% (v/v)水之非質子性溶劑中反應。 297.  如段落264至296中任一項之方法，其中步驟a)進一步包含使經碳酸衍生物活化之醣與相對於經活化醣之多醣重複單元的量(RU之莫耳當量)在0.01至10莫耳當量之間的量的疊氮基連接子反應。 298.  如段落264至297中任一項之方法，其中步驟a)之後經活化醣之活化程度在1.0至100%之間。 299   如段落264至297中任一項之方法，其中步驟a)之後經活化醣之活化程度在5至70%之間。 300.  如段落264至297中任一項之方法，其中步驟a)之後經活化醣之活化程度在15至50%之間。 301.  如段落264至297中任一項之方法，其中步驟a)之後經活化醣之活化程度在10至40%之間。 302.  如段落264至297中任一項之方法，其中步驟a)之後經活化醣之活化程度在5至15%之間。 303.  如段落264至297中任一項之方法，其中步驟a)之後經活化醣之活化程度在15至35%之間。 304.  如段落264至297中任一項之方法，其中步驟a)之後經活化醣之活化程度在15至25%之間。 305.  如段落264至297中任一項之方法，其中步驟a)之後經活化醣之活化程度為約25%。 306.  如段落264至305中任一項之方法，其中步驟b)包含使載體蛋白與相對於載體上之離胺酸0.1至10莫耳當量之量的帶有N-羥基丁二醯亞胺(NHS)部分及炔烴基之試劑反應。 307.  如段落264至306中任一項之方法，其中步驟b)之後經活化載體之活化程度在1與50之間。 308.  如段落264至306中任一項之方法，其中步驟b)之後經活化載體之活化程度可在5至50之間。 309.  如段落264至306中任一項之方法，其中步驟b)之後經活化載體之活化程度在10至40之間。 310.  如段落264至306中任一項之方法，其中步驟b)之後經活化載體之活化程度在15至25之間。 311.  如段落264至310中任一項之方法，其中結合反應c)係在水性緩衝液中進行。 312.  如段落264至310中任一項之方法，其中結合反應c)係在水性緩衝液中在作為催化劑之銅(I)存在下進行。 313.  如段落264至310中任一項之方法，其中結合反應c)係在水性緩衝液中在氧化劑及作為催化劑之銅(I)存在下進行。 314.  如段落264至313中任一項之方法，其中步驟c)處之經活化疊氮基醣與經活化炔烴-載體之初始輸入比率(重量/重量)在0.8與1.2之間。 315.  如段落264至314中任一項之方法，其中在步驟c)之後，使用適合的疊氮基封端劑對結合物中未反應之疊氮基進行封端。 316.  如段落264至315中任一項之方法，其中在步驟c)之後，使用適合的炔烴基封端劑將未反應之炔烴基進行封端。 317.  如段落283至336中任一項之方法，其中該肺炎鏈球菌血清型38醣結合物為如段落1至57中任一項的。 318.  一種使用還原胺化製備肺炎鏈球菌血清型38醣結合物之方法，該方法包含以下步驟：(1)使血清型38純化之醣氧化(活化)，(2)使經活化之醣及載體蛋白還原以形成醣結合物。 319.  如段落318之方法，其中該經分離之血清型38多醣在氧化之前經尺寸設定。 320.  一種製備肺炎鏈球菌血清型38醣結合物之方法，其包含以下步驟：(a)使該血清型38醣與氧化劑反應；(b)使步驟(a)之經活化之醣與載體蛋白混配；及(c)使混配之經活化之醣及載體蛋白與還原劑反應以形成醣結合物。 321.  如段落320之方法，其中該氧化劑為過碘酸鹽。 322.  如段落321之方法，其中該經活化之血清型38醣之氧化程度在2與30之間。 323.  如段落321之方法，其中該經活化之血清型38醣之氧化程度(DO)在10與25之間。 324.  如段落341之方法，其中該經活化之血清型38醣之氧化程度(DO)在10與25之間，且其中步驟b)處之經活化之血清型38醣與載體蛋白之初始輸入比率(重量/重量)在1.5:1與0.5:1之間。 325.  如段落321之方法，其中該經活化之血清型38醣之氧化程度(DO)在10與25之間，且其中步驟b)處之經活化之血清型38醣與載體蛋白之初始輸入比率(重量/重量)在1.5:1與0.5:1之間，其中該還原反應(c)在非質子性溶劑中進行。 326.  如段落321之方法，其中該經活化之血清型38醣之氧化程度(DO)在10與25之間，且其中步驟b)處之經活化之血清型38醣與載體蛋白之初始輸入比率(重量/重量)在1.5:1與0.5:1之間，其中該還原反應(c)在基本上由二甲亞碸(DMSO)組成之溶液中進行。 327.  如段落321之方法，其中該經活化之血清型38醣之氧化程度(DO)在10與25之間，且其中步驟b)處之經活化之血清型38醣與載體蛋白之初始輸入比率(重量/重量)在1.5:1與0.5:1之間，其中該還原反應(c)在DMSO (二甲亞碸)溶劑中進行。 328.  如段落321之方法，其中該經活化之血清型38醣之氧化程度(DO)在10與25之間，且其中步驟b)處之經活化之血清型38醣與載體蛋白之初始輸入比率(重量/重量)在1.5:1與0.5:1之間，其中該還原反應(c)在DMSO (二甲亞碸)溶劑中進行且其中該還原劑為氰基硼氫化鈉。 329.  如段落320至328中任一項之方法，其中該肺炎鏈球菌血清型38醣結合物為如段落44至74中任一項的。 330.  如段落328之方法，其中該肺炎鏈球菌血清型38醣結合物為如段落44至74中任一項的。 10 Specific embodiments of the present invention are described in the following numbered paragraphs 1 to 330 : 1. An isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: wherein n represents the number of repeating units, wherein Y represents a serine or glycine residue and wherein all of the repeating units comprise the same Y residue. 2. An isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: wherein n represents the number of repeating units. 3. An isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: wherein n represents the number of repeating units. 4. An isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: wherein n represents the number of repeating units, wherein Y represents a serine or glycine residue, wherein all of the repeating units comprise the same Y residue, and wherein the O-acetyl group at position 4 of β-D-Gal p 4OAc,6(Y) is present in about 0% to about 100% of the repeating units. 5. The isolated pneumococcal serotype 38 saccharide of paragraph 4, wherein the O-acetyl group is present in about 50% to about 100% of the repeating units. 6. The isolated pneumococcal serotype 38 saccharide of paragraph 4, wherein the O-acetyl group is present in about 80% to about 100% of the repeating units. 7. The isolated S. pneumoniae serotype 38 saccharide of paragraph 4, wherein the O-acetyl group is present in about 90% to about 100% of the repeating units. 8. An isolated S. pneumoniae serotype 38 saccharide having the following repeating units: wherein n represents the number of repeat units, wherein Y represents a serine or glycine residue, wherein all of the repeat units comprise the same Y residue, and wherein the O-acetyl group at position 4 of β-D-Gal p 4OAc,6(Y) is present in about 100% of the repeat units. 9. An isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeat units: wherein n represents the number of repeat units, wherein Y represents a serine or glycine residue, wherein all of the repeat units comprise the same Y residue, and wherein the O-acetyl group at position 4 of β-D-Gal p 4OAc,6(Y) is present in about 95% of the repeat units. 10. An isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeat units: wherein n represents the number of repeat units and wherein the O-acetyl group at position 4 of β-D-Gal p 4OAc,6Ser is present in about 0% to about 100% of the repeat units. 11. The isolated pneumococcal serotype 38 saccharide of paragraph 10, wherein the O-acetyl group is present in about 50% to about 100% of the repeat units. 12. The isolated pneumococcal serotype 38 saccharide of paragraph 10, wherein the O-acetyl group is present in about 80% to about 100% of the repeat units. 13. The isolated pneumococcal serotype 38 saccharide of paragraph 10, wherein the O-acetyl group is present in about 90% to about 100% of the repeat units. 14. An isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: wherein n represents the number of repeat units and wherein the O-acetyl group at position 4 of β-D-Gal p 4OAc,6Ser is present in about 100% of the repeat units. 15. An isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeat units: wherein n represents the number of repeat units and wherein the O-acetyl group at position 4 of β-D-Gal p 4OAc,6Ser is present in about 95% of the repeat units. 16. An isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeat units: wherein n represents the number of repeat units and wherein the O-acetyl group at position 4 of β-D-Gal p 4OAc,6Gly is present in about 0% to about 100% of the repeat units. 17. The isolated S. pneumoniae serotype 38 saccharide of paragraph 16, wherein the O-acetyl group is present in about 50% to about 100% of the repeat units. 18. The isolated S. pneumoniae serotype 38 saccharide of paragraph 16, wherein the O-acetyl group is present in about 80% to about 100% of the repeat units. 19. The isolated S. pneumoniae serotype 38 saccharide of paragraph 16, wherein the O-acetyl group is present in about 90% to about 100% of the repeat units. 20. An isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: wherein n represents the number of repeating units and wherein the O-acetyl group at position 4 of β-D-Gal p 4OAc,6Gly is present in about 100% of the repeating units. 21. An isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: wherein n represents the number of repeating units and wherein the O-acetyl group at position 4 of β-D-Gal p 4OAc,6Gly is present in 95% of the repeating units. 22. An isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: wherein n represents the number of repeating units, wherein Y represents a serine or glycine residue and wherein all of the repeating units comprise the same Y residue. 23. An isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: wherein n represents the number of repeating units. 24. An isolated Streptococcus pneumoniae serotype 38 saccharide having the following repeating units: wherein n represents the number of repeating units. 24. An isolated pneumococcal serotype 38 saccharide as described in any of paragraphs 1 to 23, having between 10 and 5,000 repeating units. 25. An isolated pneumococcal serotype 38 saccharide as described in any of paragraphs 1 to 23, having between 50 and 4,500 repeating units. 26. An isolated pneumococcal serotype 38 saccharide as described in any of paragraphs 1 to 23, having between 100 and 4,500 repeating units. 27. An isolated pneumococcal serotype 38 saccharide as described in any of paragraphs 1 to 23, having between 150 and 2,000 repeating units. 28. An isolated pneumococcal serotype 38 saccharide as described in any of paragraphs 1 to 23, having a weight average molecular weight of between 5 kDa and 5000 kDa. 29. An isolated pneumococcal serotype 38 saccharide as described in any of paragraphs 1 to 23, having a weight average molecular weight of between 5 kDa and 2000 kDa. 30. An isolated pneumococcal serotype 38 saccharide as described in any of paragraphs 1 to 23, having a weight average molecular weight of between 50 kDa and 5000 kDa. 31. An isolated pneumococcal serotype 38 saccharide as described in any of paragraphs 1 to 23, having a weight average molecular weight of between 50 kDa and 1000 kDa. 32. An isolated pneumococcal serotype 38 saccharide as described in any of paragraphs 1 to 23, having a weight average molecular weight of between 100 kDa and 5000 kDa. 33. An isolated pneumococcal serotype 38 saccharide as described in any of paragraphs 1 to 23, having a weight average molecular weight of between 100 kDa and 1000 kDa. 34. An isolated pneumococcal serotype 38 saccharide as described in any of paragraphs 1 to 23, having a weight average molecular weight of between 100 kDa and 500 kDa. 35. An isolated pneumococcal serotype 38 saccharide as described in any of paragraphs 1 to 23, having a weight average molecular weight of between 300 kDa and 5000 kDa. 36. An isolated pneumococcal serotype 38 saccharide as described in any of paragraphs 1 to 23, having a weight average molecular weight of between 300 kDa and 1000 kDa. 37. An isolated pneumococcal serotype 38 saccharide as described in any of paragraphs 1 to 23, having a weight average molecular weight of between 500 kDa and 3000 kDa. 38. An isolated pneumococcal serotype 38 saccharide as described in any of paragraphs 1 to 23, having a weight average molecular weight of between 500 kDa and 2000 kDa. 39. An isolated pneumococcal serotype 38 saccharide as described in any of paragraphs 1 to 23, having a weight average molecular weight of between 500 kDa and 1000 kDa. 40. An isolated pneumococcal serotype 38 saccharide as described in any of paragraphs 1 to 23, having a size set to a weight average molecular weight between 10 kDa and 1000 kDa. 41. An isolated pneumococcal serotype 38 saccharide as described in any of paragraphs 1 to 23, having a size set to a weight average molecular weight between 50 kDa and 500 kDa. 42. An isolated pneumococcal serotype 38 saccharide as described in any of paragraphs 1 to 23, having a size set to a weight average molecular weight between 100 kDa and 400 kDa. 43. A glycoconjugate comprising an isolated pneumococcal serotype 38 saccharide as described in any of paragraphs 1 to 42 conjugated to a carrier protein. 44. A glycoconjugate consisting of an isolated Streptococcus pneumoniae serotype 38 saccharide as described in any one of paragraphs 1 to 42 conjugated to a carrier protein. 45. A serotype 38 saccharide conjugate comprising a serotype 38 capsular saccharide, which contains about 1 to about 70 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide. 46. The serotype 38 saccharide conjugate of paragraph 45, which contains about 1 to about 68 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide. 46. The serotype 38 glycoconjugate of paragraph 45, comprising about 1 to about 65 N-acetyl-D-fucosamine (D-FucNAc) residues per 100 carbohydrate repeat units of the saccharide. 47. A serotype 38 glycoconjugate comprising serotype 38 capsular saccharide, comprising about 0.5 to about 35 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide. 48. The serotype 38 glycoconjugate of paragraph 47, comprising about 0.5 to about 32 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide. 49. The serotype 38 glycoconjugate of paragraph 47, comprising about 0.5 to about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide. 50. A serotype 38 glycoconjugate comprising a serotype 38 capsular saccharide, comprising about 1 to about 70 N-acetyl-D-fucosamine (D-FucNAc) residues and about 0.5 to about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide. 51. A serotype 38 glycoconjugate comprising a serotype 38 capsular saccharide comprising about 1 to about 68 N-acetyl-D-fucosamine (D-FucNAc) residues and about 0.5 to about 32 N-acetyl-D-quinolfuramine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide. 52. A serotype 38 glycoconjugate comprising a serotype 38 capsular saccharide comprising about 1 to about 65 N-acetyl-D-fucosamine (D-FucNAc) residues and about 0.5 to about 32.5 N-acetyl-D-quinolfuramine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide. 53. A serotype 38 glycoconjugate comprising a serotype 38 capsular saccharide comprising about 1 to about 60 N-acetyl-D-fucosamine (D-FucNAc) residues and about 0.5 to about 30 N-acetyl-D-quinolfuramine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide. 54. A serotype 38 glycoconjugate comprising a serotype 38 capsular saccharide comprising about 68 N-acetyl-D-fucosamine (D-FucNAc) residues and about 32 N-acetyl-D-quinolfuramine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide. 55. A serotype 38 glycoconjugate according to any one of paragraphs 45 to 54, comprising serotype 38 capsular saccharide comprising N-acetyl-D-fucosamine (D-FucNAc) residues and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues, wherein the number of D-FucNAc residues is about twice the number of D-QuiNAc residues. 56. A serotype 38 glycoconjugate comprising serotype 38 capsular saccharide comprising N-acetyl-D-fucosamine (D-FucNAc), N-acetyl-D-quinolfuramine (D-QuiNAc) and D-2-acetamido-2,6-dideoxy-xylose-hex-4-ketose (D-Sug) residues. 57. A serotype 38 glycoconjugate as described in paragraph 56, comprising a serotype 38 glycoconjugate comprising an N-acetyl-D-fucosamine (D-FucNAc) residue, an N-acetyl-D-quinolfuramine (D-QuiNAc) residue, or a D-2-acetamido-2,6-dideoxy-xylose-hex-4-ketose (D-Sug) residue at the same position of the repeat unit. 58. A serotype 38 glycoconjugate as described in paragraph 56 or 57, wherein the number of D-FucNAc residues is about twice the number of D-QuiNAc residues. 59. A serotype 38 glycoconjugate comprising a serotype 38 glycoconjugate having the following repeat units: wherein n represents the number of repeat units, wherein Y represents a serine or glycine residue, wherein all such repeat units contain the same Y residue, and wherein X represents an N-acetyl-D-fucosamine (D-FucNAc) residue or an N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue. 60. A serotype 38 glycoconjugate as described in paragraph 59, wherein the serotype 38 capsane contains about 70 N-acetyl-D-fucosamine (D-FucNAc) residues and about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide. 61. The serotype 38 glycoconjugate of paragraph 59, wherein the serotype 38 capsular saccharide comprises about 68 N-acetyl-D-fucosamine (D-FucNAc) residues and about 32 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide. 62. A serotype 38 glycoconjugate comprising a serotype 38 capsular saccharide having the following repeat units: wherein n represents the number of repeat units, wherein Y represents a serine or glycine residue, wherein all such repeat units contain the same Y residue, and wherein X represents an N-acetyl-D-fucosamine (D-FucNAc) residue or an N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue. 63. A serotype 38 glycoconjugate as described in paragraph 62, wherein the serotype 38 capsane contains about 70 N-acetyl-D-fucosamine (D-FucNAc) residues and about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide. 64. The serotype 38 glycoconjugate of paragraph 62, wherein the serotype 38 capsular saccharide comprises about 68 N-acetyl-D-fucosamine (D-FucNAc) residues and about 32 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide. 65. A serotype 38 glycoconjugate comprising a serotype 38 capsular saccharide having the following repeat units: wherein n represents the number of repeating units and wherein X represents an N-acetyl-D-fucosamine (D-FucNAc) residue, an N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue or a D-2-acetamido-2,6-dideoxy-xylose-hex-4-ketose (D-Sug) residue. 66. The serotype 38 glycoconjugate of paragraph 65, wherein the serotype 38 capsular saccharide comprises about 1 to about 70 N-acetyl-D-fucosamine (D-FucNAc) residues, about 0.5 to about 30 N-acetyl-D-quinolfuramine (D-QuiNAc) residues, and about 0 to about 98.5 D-2-acetamido-2,6-dideoxy-xylose-hex-4-ketose (D-Sug) residues per 100 carbohydrate repeat units of the saccharide. 67. A serotype 38 glycoconjugate comprising a serotype 38 capsular saccharide having the following repeat units: wherein n represents the number of repeat units and wherein X represents an N-acetyl-D-fucosamine (D-FucNAc) residue or an N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue. 68. The serotype 38 glycoconjugate of paragraph 67, wherein the serotype 38 capsular saccharide comprises about 70 N-acetyl-D-fucosamine (D-FucNAc) residues and about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide. 69. The serotype 38 glycoconjugate of paragraph 67, wherein the serotype 38 capsular saccharide comprises about 68 N-acetyl-D-fucosamine (D-FucNAc) residues and about 32 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide. 70. A serotype 38 glycoconjugate comprising a serotype 38 capsular saccharide having the following repeat units: wherein n represents the number of repeating units and wherein X represents an N-acetyl-D-fucosamine (D-FucNAc) residue, an N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue or a D-2-acetamido-2,6-dideoxy-xylose-hex-4-ketose (D-Sug) residue. 71. The serotype 38 glycoconjugate of paragraph 70, wherein the serotype 38 capsular saccharide comprises about 1 to about 70 N-acetyl-D-fucosamine (D-FucNAc) residues, about 0.5 to about 30 N-acetyl-D-quinolfuramine (D-QuiNAc) residues, and about 0 to about 98.5 D-2-acetamido-2,6-dideoxy-xylose-hex-4-ketose (D-Sug) residues per 100 carbohydrate repeat units of the saccharide. 72. A serotype 38 glycoconjugate comprising a serotype 38 capsular saccharide having the following repeat units: wherein n represents the number of repeat units and wherein X represents an N-acetyl-D-fucosamine (D-FucNAc) residue or an N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue. 73. The serotype 38 glycoconjugate of paragraph 72, wherein the serotype 38 capsular saccharide comprises about 70 N-acetyl-D-fucosamine (D-FucNAc) residues and about 30 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide. 74. The serotype 38 glycoconjugate of paragraph 72, wherein the serotype 38 capsular saccharide comprises about 68 N-acetyl-D-fucosamine (D-FucNAc) residues and about 32 N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues per 100 carbohydrate repeat units of the saccharide. 75. The serotype 38 glycoconjugate of paragraphs 43 or 44, wherein the serotype 38 glycoconjugate comprises a serotype 38 saccharide covalently bound to a carrier protein via an eTEC spacer. 75. The serotype 38 glycoconjugate of paragraphs 43 or 44, wherein the serotype 38 glycoconjugate is prepared by click chemistry. 76. A serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (IV): , wherein X is selected from the group consisting of CH 2 (CH 2 ) n ′ , (CH 2 CH 2 O) m CH 2 CH 2 , NHCO(CH 2 ) n ′ , NHCO(CH 2 CH 2 O) m CH 2 CH 2 , OCH 2 (CH 2 ) n ′ , and O(CH 2 CH 2 O) m CH 2 CH 2 ; wherein n′ is selected from 1 to 10 and m is selected from 1 to 4, wherein X′ is selected from the group consisting of CH 2 O(CH 2 ) n′ CH 2 C═O, CH 2 O(CH 2 CH 2 O) m′ (CH 2 ) n′ CH 2 C═O, wherein n′ is selected from 0 to 10 and m′ is selected from 0 to 4, wherein the structure in square brackets represents the repeating unit of the serotype 38 carbohydrate, and wherein n represents the number of repeating units. 77. A serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (IV), wherein X is CH 2 (CH 2 ) n ′ , wherein n′ is 2, and wherein X′ is CH 2 O(CH 2 ) n ′′ CH 2 C═O, wherein n′′ is 1. 78. A serotype 38 saccharide conjugate, comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (V), wherein the structure in square brackets represents the repeating unit of the serotype 38 saccharide, and wherein n represents the number of repeating units. 79. A serotype 38 saccharide conjugate comprising a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and having the general formula (XII): wherein X is selected from the group consisting of CH2(CH2)n', (CH2CH2O ) mCH2CH2 , NHCO ( CH2 ) n ' , NHCO ( CH2CH2O ) mCH2CH2 , OCH2 ( CH2 ) n ' , and O( CH2CH2O ) mCH2CH2 ; wherein n' is selected from 0 to 10 and m is selected from 1 to 4, wherein X' is selected from the group consisting of CH2 ( CH2 ) n '' , CH2O ( CH2 ) n''CH2 , CH2O (CH2CH2O)m ' , ( CH2 ) n''CH 2 , wherein n'' is selected from 0 to 10 and m' is selected from 0 to 4, and wherein the structure in square brackets represents the repeating unit of the serotype 38 saccharide, and wherein n represents the number of repeating units. 80. A serotype 38 saccharide conjugate, which comprises a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and has the general formula (IV), wherein X is CH 2 (CH 2 ) n ' , wherein n' is 0, and wherein X' is CH 2 (CH 2 ) n '' , wherein n'' is 0. 81. A serotype 38 saccharide conjugate, which comprises a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and has the general formula (XIII), wherein the structure in square brackets represents a repeating unit of the serotype 38 saccharide, and wherein n represents the number of repeating units. 80. The glycoconjugate of any of paragraphs 45 to 81, wherein the serotype 38 saccharide is an isolated Streptococcus pneumoniae serotype 38 saccharide of any of paragraphs 1 to 42. 81. The glycoconjugate of any of paragraphs 43 to 81, wherein the weight average molecular weight (Mw) of the Streptococcus pneumoniae serotype 38 saccharide prior to conjugation is between 50 kDa and 1,000 kDa. 82. The glycoconjugate of any of paragraphs 43 to 81, wherein the weight average molecular weight (Mw) of the Streptococcus pneumoniae serotype 38 saccharide prior to conjugation is between 100 kDa and 600 kDa. 83. The glycoconjugate of any of paragraphs 43 to 81, wherein the weight average molecular weight (Mw) of the pneumococcal serotype 38 saccharide prior to conjugation is between 100 kDa and 400 kDa. 84. The glycoconjugate of any of paragraphs 43 to 81, wherein the weight average molecular weight (Mw) of the pneumococcal serotype 38 saccharide prior to conjugation is between 150 kDa and 300 kDa. 85. The glycoconjugate of any of paragraphs 43 to 84, wherein the weight average molecular weight (Mw) of the serotype 38 saccharide is between 250 kDa and 20,000 kDa. 86. The glycoconjugate of any of paragraphs 43 to 84, wherein the weight average molecular weight (Mw) of the serotype 38 saccharide is between 500 kDa and 15,000 kDa. 87. The glycoconjugate of any of paragraphs 43 to 84, wherein the weight average molecular weight (Mw) of the serotype 38 glycoconjugate is between 500 kDa and 10,000 kDa. 87a. The glycoconjugate of any of paragraphs 43 to 84, wherein the weight average molecular weight (Mw) of the serotype 38 glycoconjugate is between 1,000 kDa and 10,000 kDa. 87b. The glycoconjugate of any of paragraphs 43 to 81, wherein the weight average molecular weight (Mw) of the Streptococcus pneumoniae serotype 38 saccharide prior to conjugation is between 150 kDa and 300 kDa, and wherein the weight average molecular weight (Mw) of the serotype 38 glycoconjugate is between 1,000 kDa and 10,000 kDa. 88. The glycoconjugate of any of paragraphs 43 to 84, wherein the weight average molecular weight (Mw) of the serotype 38 glycoconjugate is between 750 kDa and 7,500 kDa. 89. The glycoconjugate of any of paragraphs 43 to 84, wherein the weight average molecular weight (Mw) of the serotype 38 glycoconjugate is between 1,000 kDa and 5,000 kDa. 90. The glycoconjugate of any of paragraphs 43 to 89, wherein the degree of conjugation of the serotype 38 glycoconjugate is between 2 and 15. 91. The glycoconjugate of any of paragraphs 43 to 89, wherein the degree of conjugation of the serotype 38 glycoconjugate is between 2 and 10. 92. The glycoconjugate of any of paragraphs 43 to 89, wherein the degree of binding of the serotype 38 glycoconjugate is between 3 and 5. 93. The glycoconjugate of any of paragraphs 43 to 89, wherein the degree of binding of the serotype 38 glycoconjugate is between 2 and 6. 94. The glycoconjugate of any of paragraphs 43 to 89, wherein the degree of binding of the serotype 38 glycoconjugate is between 4 and 10. 94a. The glycoconjugate of any of paragraphs 43 to 81, wherein the weight average molecular weight (Mw) of the Streptococcus pneumoniae serotype 38 saccharide prior to conjugation is between 150 kDa and 300 kDa, wherein the weight average molecular weight (Mw) of the serotype 38 glycoconjugate is between 1,000 kDa and 10,000 kDa and wherein the degree of conjugation of the serotype 38 glycoconjugate is between 2 and 10. 94b. The glycoconjugate of any of paragraphs 43 to 81, wherein the weight average molecular weight (Mw) of the pneumococcal serotype 38 saccharide prior to conjugation is between 150 kDa and 300 kDa, wherein the weight average molecular weight (Mw) of the serotype 38 saccharide conjugate is between 1,000 kDa and 10,000 kDa and wherein the degree of conjugation of the serotype 38 saccharide conjugate is between 4 and 10. 95. The glycoconjugate of any of paragraphs 43 to 94, wherein the ratio (w/w) of serotype 38 saccharide to carrier protein in the glycoconjugate is between 0.5 and 3.0. 97. The glycoconjugate of any of paragraphs 43 to 94, wherein the ratio (w/w) of serotype 38 saccharide to carrier protein in the glycoconjugate is between 0.5 and 2.0. 98. The glycoconjugate of any one of paragraphs 43 to 94, wherein the ratio (w/w) of serotype 38 saccharides to carrier protein in the glycoconjugate is between 0.5 and 1.5. 99. The glycoconjugate of any one of paragraphs 43 to 94, wherein the ratio (w/w) of serotype 38 saccharides to carrier protein in the glycoconjugate is between 0.8 and 1.2. 100. The glycoconjugate of any one of paragraphs 43 to 94, wherein the ratio (w/w) of serotype 38 saccharides to carrier protein in the glycoconjugate is between 0.5 and 1.0. 101. The glycoconjugate of any one of paragraphs 43 to 94, wherein the ratio (w/w) of serotype 38 saccharides to carrier protein in the glycoconjugate is between 1.0 and 1.5. 102. The glycoconjugate of any one of paragraphs 43 to 94, wherein the ratio of serotype 38 saccharides to carrier protein (w/w) in the glycoconjugate is between 1.0 and 2.0. 103. The glycoconjugate of any one of paragraphs 43 to 94, wherein the ratio of serotype 38 saccharides to carrier protein (w/w) in the glycoconjugate is between 0.8 and 1.2. 104. The glycoconjugate of any one of paragraphs 43 to 94, wherein the ratio of serotype 38 saccharides to carrier protein (w/w) in the glycoconjugate is between 0.7 and 1.1. 104a. The glycoconjugate of any of paragraphs 43 to 81, wherein the weight average molecular weight (Mw) of the Streptococcus pneumoniae serotype 38 saccharide prior to conjugation is between 150 kDa and 300 kDa, wherein the weight average molecular weight (Mw) of the serotype 38 glycoconjugate is between 1,000 kDa and 10,000 kDa, wherein the ratio (w/w) of serotype 38 saccharide to carrier protein in the glycoconjugate is between 0.5 and 1.5 and wherein the degree of conjugation of the serotype 38 glycoconjugate is between 4 and 10. 104b. The glycoconjugate of any of paragraphs 43 to 81, wherein the weight average molecular weight (Mw) of the S. pneumoniae serotype 38 saccharide prior to conjugation is between 150 kDa and 300 kDa, wherein the weight average molecular weight (Mw) of the serotype 38 saccharide conjugate is between 1,000 kDa and 10,000 kDa, wherein the ratio (w/w) of serotype 38 saccharide to carrier protein in the glycoconjugate is between 0.7 and 1.1 and wherein the degree of conjugation of the serotype 38 saccharide conjugate is between 4 and 10. 105. The glycoconjugate of any of paragraphs 43 to 104, wherein the serotype 38 saccharide conjugate comprises less than about 50% free serotype 38 saccharide compared to the total amount of serotype 38 saccharide. 106. The glycoconjugate of any of paragraphs 43 to 104, wherein the serotype 38 glycoconjugate comprises less than about 25% free serotype 38 saccharides compared to the total amount of serotype 38 saccharides. 107. The glycoconjugate of any of paragraphs 43 to 104, wherein the serotype 38 glycoconjugate comprises less than about 20% free serotype 38 saccharides compared to the total amount of serotype 38 saccharides. 108. The glycoconjugate of any of paragraphs 43 to 104, wherein the serotype 38 glycoconjugate comprises less than about 15% free serotype 38 saccharides compared to the total amount of serotype 38 saccharides. 108a. The glycoconjugate of any of paragraphs 43 to 81, wherein the weight average molecular weight (Mw) of the Streptococcus pneumoniae serotype 38 saccharide prior to conjugation is between 150 kDa and 300 kDa, wherein the weight average molecular weight (Mw) of the serotype 38 saccharide conjugate is between 1,000 kDa and 10,000 kDa, wherein the ratio (w/w) of serotype 38 saccharide to carrier protein in the glycoconjugate is between 0.5 and 1.5, wherein the degree of conjugation of the serotype 38 saccharide conjugate is between 4 and 10 and wherein the serotype 38 saccharide conjugate comprises less than about 20% free serotype 38 saccharide compared to the total amount of serotype 38 saccharide. 108b. The glycoconjugate of any of paragraphs 43 to 81, wherein the weight average molecular weight (Mw) of the Streptococcus pneumoniae serotype 38 saccharide prior to conjugation is between 150 kDa and 300 kDa, wherein the weight average molecular weight (Mw) of the serotype 38 saccharide conjugate is between 1,000 kDa and 10,000 kDa, wherein the ratio (w/w) of serotype 38 saccharide to carrier protein in the glycoconjugate is between 0.7 and 1.1, wherein the degree of conjugation of the serotype 38 saccharide conjugate is between 4 and 10 and wherein the serotype 38 saccharide conjugate comprises less than about 20% free serotype 38 saccharide compared to the total amount of serotype 38 saccharide. 109. The glycoconjugate of any of paragraphs 43 to 108, wherein at least 30% of the serotype 38 glycoconjugate has a K d of less than or equal to 0.3 in a CL-4B column. 110. The glycoconjugate of any of paragraphs 43 to 108, wherein at least 40% of the serotype 38 glycoconjugate has a K d of less than or equal to 0.3 in a CL-4B column. 111. The glycoconjugate of any of paragraphs 43 to 108, wherein at least 60% of the serotype 38 glycoconjugate has a K d of less than or equal to 0.3 in a CL-4B column. 112. The glycoconjugate of any of paragraphs 43 to 108, wherein between 50% and 80% of the serotype 38 glycoconjugate has a K d of less than or equal to 0.3 in a CL-4B column. 113. The glycoconjugate of any one of paragraphs 43 to 108, wherein between 65% and 80% of the serotype 38 glycoconjugate has a K d less than or equal to 0.3 in a CL-4B column. 113a. The glycoconjugate of any of paragraphs 43 to 81, wherein the weight average molecular weight (Mw) of the Streptococcus pneumoniae serotype 38 saccharide prior to conjugation is between 150 kDa and 300 kDa, wherein the weight average molecular weight (Mw) of the serotype 38 saccharide conjugate is between 1,000 kDa and 10,000 kDa, wherein the ratio (w/w) of serotype 38 saccharide to carrier protein in the glycoconjugate is between 0.5 and 1.5, wherein the degree of conjugation of the serotype 38 saccharide conjugate is between 4 and 10, wherein the serotype 38 saccharide conjugate comprises less than about 20% free serotype 38 saccharide compared to the total amount of serotype 38 saccharide and wherein between 65% and 80% of the serotype 38 saccharide conjugate is bound to the carrier protein at 40 °C in the CL-4B column. d is less than or equal to 0.3. 113b. The glycoconjugate of any of paragraphs 43 to 81, wherein the weight average molecular weight (Mw) of the Streptococcus pneumoniae serotype 38 saccharide prior to conjugation is between 150 kDa and 300 kDa, wherein the weight average molecular weight (Mw) of the serotype 38 saccharide conjugate is between 1,000 kDa and 10,000 kDa, wherein the ratio (w/w) of serotype 38 saccharide to carrier protein in the glycoconjugate is between 0.7 and 1.1, wherein the degree of conjugation of the serotype 38 saccharide conjugate is between 4 and 10, wherein the serotype 38 saccharide conjugate comprises less than about 20% free serotype 38 saccharide compared to the total amount of serotype 38 saccharide and wherein between 65% and 80% of the serotype 38 saccharide conjugate is bound to the carrier protein at 40°C in the CL-4B column. d is less than or equal to 0.3. 114. The glycoconjugate of any of paragraphs 43 to 113, wherein the carrier protein of the serotype 38 glycoconjugate is selected from the group consisting of TT, DT, DT mutants and C5a peptidase (SCP) from Streptococcus. 115. The glycoconjugate of any of paragraphs 43 to 113, wherein the carrier protein of the serotype 38 glycoconjugate is Rhizobium avidin [aa 45-179J-GGGGSSS-SP1500- AAA-SP0785] (CP1). 115a. The glycoconjugate of any of paragraphs 43 to 113, wherein the carrier protein of the serotype 38 glycoconjugate is Rhavi-linker-PdT(G294P)-linker-SP0435 [aa 62-185] fusion protein (SPP2). 116. The glycoconjugate of any of paragraphs 43 to 113, wherein the carrier protein of the serotype 38 glycoconjugate is DT. 117. The glycoconjugate of any of paragraphs 43 to 113, wherein the carrier protein of the serotype 38 glycoconjugate is TT. 118. The glycoconjugate of any of paragraphs 43 to 113, wherein the carrier protein of the serotype 38 glycoconjugate is PD. 119. The glycoconjugate of any of paragraphs 43 to 113, wherein the carrier protein of the serotype 38 glycoconjugate is CRM 197 or C5a peptidase (SCP) from Streptococcus. 120. The glycoconjugate of any of paragraphs 43 to 113, wherein the carrier protein of the serotype 38 glycoconjugate is CRM 197. 121. The glycoconjugate of any of paragraphs 43 to 113, wherein the carrier protein of the serotype 38 glycoconjugate is SCP. 122. The glycoconjugate of any of paragraphs 43 to 113, wherein the carrier protein of the serotype 38 glycoconjugate is enzymatically inactive SCP. 123. The glycoconjugate of any of paragraphs 43 to 113, wherein the carrier protein of the serotype 38 glycoconjugate is an enzymatically inactive SCP from GBS (SCPB). 124. The glycoconjugate of any of paragraphs 43 to 113, wherein the carrier protein of the serotype 38 glycoconjugate is an enzymatically inactive SCP from GAS (SCPA). 125. The glycoconjugate of any of paragraphs 43 to 113, wherein the carrier protein of the serotype 38 glycoconjugate is a fragment of SCPB. 126. The glycoconjugate of any of paragraphs 43 to 113, wherein the carrier protein of the serotype 38 glycoconjugate is an enzymatically inactive fragment of SCP consisting of SEQ ID NO: 1. 127. The glycoconjugate of any of paragraphs 43 to 113, wherein the carrier protein of the serotype 38 glycoconjugate is an enzymatically inactive fragment of SCP consisting of SEQ ID NO: 2. 128. An immunogenic composition comprising the pneumococcal serotype 38 saccharide of any of paragraphs 1 to 42. 129. An immunogenic composition comprising the pneumococcal serotype 38 glycoconjugate of any of paragraphs 43 to 127. 130. The immunogenic composition of paragraph 129, comprising 1 to 45 different glycoconjugates. 131. The immunogenic composition of paragraph 129, comprising glycoconjugates from 1 to 45 different pneumococcal serotypes. 132. The immunogenic composition of paragraph 129, comprising saccharide conjugates from 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 different serotypes of Streptococcus pneumoniae. 133. The immunogenic composition of paragraph 129, comprising saccharide conjugates from 16 or 20 different serotypes of Streptococcus pneumoniae. 134. The immunogenic composition of paragraph 129, which is a 21, 22, 23, 24 or 25 valent Streptococcus pneumoniae conjugate composition. 135. The immunogenic composition of paragraph 129, which is a 21 valent Streptococcus pneumoniae conjugate composition. 136. The immunogenic composition of paragraph 129, which is a 22 valent Streptococcus pneumoniae conjugate composition. 137. The immunogenic composition of paragraph 129, which is a 23-valent pneumococcal conjugate composition. 138. The immunogenic composition of paragraph 129, which is a 24-valent pneumococcal conjugate composition. 139. The immunogenic composition of paragraph 129, which is a 25-valent pneumococcal conjugate composition. 140. The immunogenic composition of paragraph 129, which comprises glycoconjugates from 26 to 45 different pneumococcal serotypes. 141. The immunogenic composition of paragraph 129, which comprises glycoconjugates from 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44 or 45 different pneumococcal serotypes. 142. The immunogenic composition of paragraph 129, comprising saccharide conjugates from 35 or 45 different serotypes of Streptococcus pneumoniae. 142a. The immunogenic composition of paragraph 129, comprising saccharide conjugates from 35 different serotypes of Streptococcus pneumoniae. 143. The immunogenic composition of paragraph 129, which is a 35-, 36-, 37-, 38-, 39-, 40-, 41-, 42-, 43-, 44-, or 45-valent Streptococcus pneumoniae conjugate composition. 143a. The immunogenic composition of paragraph 129, which is a 35-valent Streptococcus pneumoniae conjugate composition. 144. The immunogenic composition of paragraph 129, which is a 40-, 41-, 42-, 43-, 44-, or 45-valent Streptococcus pneumoniae conjugate composition. 145. The immunogenic composition of paragraph 129, which is a 40-valent pneumococcal conjugate composition. 146. The immunogenic composition of paragraph 129, which is a 41-valent pneumococcal conjugate composition. 147. The immunogenic composition of paragraph 129, which is a 42-valent pneumococcal conjugate composition. 148. The immunogenic composition of paragraph 129, which is a 43-valent pneumococcal conjugate composition. 149. The immunogenic composition of paragraph 129, which is a 44-valent pneumococcal conjugate composition. 150. The immunogenic composition of paragraph 129, which is a 45-valent pneumococcal conjugate composition. 151. The immunogenic composition of any of paragraphs 129 to 150, further comprising saccharide conjugates from pneumococcal serotypes 4, 6B, 9V, 14, 18C, 19F and 23F. 152. The immunogenic composition of paragraph 151, further comprising saccharide conjugates from pneumococcal serotypes 1, 5 and 7F. 153. The immunogenic composition of paragraph 152, further comprising saccharide conjugates from pneumococcal serotype 3. 154. The immunogenic composition of paragraph 153, further comprising saccharide conjugates from pneumococcal serotypes 6A and 19A. 155. The immunogenic composition of paragraph 154, further comprising saccharide conjugates from pneumococcal serotypes 22F and 33F. 156. The immunogenic composition of paragraph 155, further comprising a glycoconjugate from pneumococcal serotypes 8, 10A, 11A, 12F and 15B. 157. The immunogenic composition of paragraph 156, further comprising a glycoconjugate from pneumococcal serotype 2. 158. The immunogenic composition of paragraph 157, further comprising a glycoconjugate from pneumococcal serotype 9N. 159. The immunogenic composition of paragraph 158, further comprising a glycoconjugate from pneumococcal serotype 17F. 160. The immunogenic composition of paragraph 159, further comprising a glycoconjugate from pneumococcal serotype 20. 161. The immunogenic composition of paragraph 160, further comprising a saccharide conjugate from Streptococcus pneumoniae serotype 2. 162. The immunogenic composition of paragraph 161, further comprising a saccharide conjugate from Streptococcus pneumoniae serotype 15C. 163. The immunogenic composition of any of paragraphs 129 to 150, further comprising a saccharide conjugate from Streptococcus pneumoniae serotypes 4, 6B, 9V, 14, 18C, 19F and 23F, and wherein the immunogenic composition is an 8-valent Streptococcus pneumoniae conjugate composition. 164. The immunogenic composition of any one of paragraphs 129 to 150, further comprising saccharide conjugates from pneumococcal serotypes 1, 4, 5, 6B, 7F, 9V, 14, 18C, 19F and 23F, and wherein the immunogenic composition is an 11-valent pneumococcal conjugate composition. 165. The immunogenic composition of any one of paragraphs 129 to 150, further comprising saccharide conjugates from pneumococcal serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F and 23F, and wherein the immunogenic composition is a 14-valent pneumococcal conjugate composition. 166. The immunogenic composition of any of paragraphs 129 to 150, further comprising saccharide conjugates from pneumococcal serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, 22F, 23F and 33F, and wherein the immunogenic composition is a 16-valent pneumococcal conjugate composition. 167. The immunogenic composition of any of paragraphs 129 to 150, further comprising saccharide conjugates from pneumococcal serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F, and wherein the immunogenic composition is a 21-valent pneumococcal conjugate composition. 168. The immunogenic composition of any one of paragraphs 129 to 150, further comprising a saccharide conjugate from Streptococcus pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15C, 18C, 19A, 19F, 22F, 23F and 33F. 169. The immunogenic composition of paragraph 168, which is a 21-valent Streptococcus pneumoniae conjugate composition. 170. The immunogenic composition of any one of paragraphs 129 to 150, further comprising a saccharide conjugate from Streptococcus pneumoniae serotypes 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F and 33F. 171. The immunogenic composition of paragraph 170, which is a 22-valent Streptococcus pneumoniae conjugate composition. 172. The immunogenic composition of any one of paragraphs 129 to 150, further comprising a saccharide conjugate from Streptococcus pneumoniae serotypes 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15C, 18C, 19A, 19F, 22F, 23F and 33F. 173. The immunogenic composition of paragraph 172, which is a 22-valent Streptococcus pneumoniae conjugate composition. 174. The immunogenic composition of any one of paragraphs 129 to 150, further comprising a saccharide conjugate from Streptococcus pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 9N, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F and 33F. 175. The immunogenic composition of paragraph 174, which is a 22-valent Streptococcus pneumoniae conjugate composition. 176. The immunogenic composition of any one of paragraphs 129 to 150, further comprising a saccharide conjugate from Streptococcus pneumoniae serotypes 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 9N, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F and 33F. 177. The immunogenic composition of paragraph 176, which is a 23-valent Streptococcus pneumoniae conjugate composition. 178. The immunogenic composition of any one of paragraphs 129 to 150, further comprising a saccharide conjugate from Streptococcus pneumoniae serotypes 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 9N, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 22F, 23F and 33F. 179. The immunogenic composition of paragraph 178, which is a 24-valent Streptococcus pneumoniae conjugate composition. 180. The immunogenic composition of any one of paragraphs 129 to 150, further comprising a saccharide conjugate from Streptococcus pneumoniae serotypes 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 9N, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F and 33F. 181. The immunogenic composition of paragraph 180, which is a 25-valent Streptococcus pneumoniae conjugate composition. 182. The immunogenic composition of any one of paragraphs 129 to 150, further comprising a saccharide conjugate from Streptococcus pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23F and 33F. 183. The immunogenic composition of paragraph 182, which is a 22-valent Streptococcus pneumoniae conjugate composition. 184. The immunogenic composition of any one of paragraphs 129 to 150, further comprising a saccharide conjugate from Streptococcus pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23A, 23F and 33F. 185. The immunogenic composition of paragraph 184, which is a 22-valent Streptococcus pneumoniae conjugate composition. 186. The immunogenic composition of any one of paragraphs 129 to 150, further comprising a saccharide conjugate from Streptococcus pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23B, 23F and 33F. 187. The immunogenic composition of paragraph 186, which is a 22-valent Streptococcus pneumoniae conjugate composition. 188. The immunogenic composition of any one of paragraphs 129 to 150, further comprising a saccharide conjugate from Streptococcus pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F, 24F and 33F. 189. The immunogenic composition of paragraph 188, which is a 22-valent Streptococcus pneumoniae conjugate composition. 190. The immunogenic composition of any one of paragraphs 129 to 150, further comprising a saccharide conjugate from Streptococcus pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F, 33F and 35B. 191. The immunogenic composition of paragraph 190, which is a 22-valent Streptococcus pneumoniae conjugate composition. 192. The immunogenic composition of any one of paragraphs 129 to 150, further comprising a saccharide conjugate from Streptococcus pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23A, 23F and 33F. 193. The immunogenic composition of paragraph 192, which is a 23-valent Streptococcus pneumoniae conjugate composition. 194. The immunogenic composition of any one of paragraphs 129 to 150, further comprising a saccharide conjugate from Streptococcus pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F and 35B. 195. The immunogenic composition of paragraph 194, wherein the Streptococcus pneumoniae saccharide is conjugated to CRM 197 . 196. The immunogenic composition of paragraph 194, wherein the pneumococcal saccharides from serotypes 1, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, 35B and 38 are conjugated to CRM 197 and the pneumococcal saccharide serotype 3 is conjugated to SCP. 197. The immunogenic composition of paragraph 196, which is a 26-valent pneumococcal conjugate composition. 198. The immunogenic composition of any of paragraphs 129 to 150, further comprising at least one saccharide conjugate selected from the group consisting of saccharide conjugates from Streptococcus pneumoniae serotypes 2, 3, 7C, 9N, 10B, 15A, 16F, 17F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B and 35F. 199. The immunogenic composition of any one of paragraphs 129 to 150, further comprising twenty-one saccharide conjugates selected from the group consisting of saccharide conjugates from Streptococcus pneumoniae serotypes 2, 3, 7C, 9N, 10B, 15A, 16F, 17F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B and 35F. 200. The immunogenic composition of paragraph 199, which is a 22-valent Streptococcus pneumoniae conjugate composition. 201. The immunogenic composition of any one of paragraphs 129 to 150, further comprising a carbohydrate conjugate from serotypes 2, 3, 7C, 9N, 10B, 15A, 16F, 17F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B and 35. 202. The immunogenic composition of paragraph 201, which is a 23-valent Streptococcus pneumoniae conjugate composition. 203. The immunogenic composition of any of paragraphs 129 to 150, further comprising at least one saccharide conjugate selected from the group consisting of saccharide conjugates from Streptococcus pneumoniae serotypes 2, 3, 7C, 9N, 10B, 15A, 16F, 17F, 19A, 19F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B and 35F. 204. The immunogenic composition of any one of paragraphs 129 to 150, further comprising twenty-two saccharide conjugates selected from the group consisting of saccharide conjugates from Streptococcus pneumoniae serotypes 2, 3, 7C, 9N, 10B, 15A, 16F, 17F, 19A, 19F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B and 35F. 205. The immunogenic composition of paragraph 204, which is a 23-valent Streptococcus pneumoniae conjugate composition. 206. The immunogenic composition of any one of paragraphs 129 to 150, further comprising twenty-three saccharide conjugates selected from the group consisting of saccharide conjugates from Streptococcus pneumoniae serotypes 2, 3, 7C, 9N, 10B, 15A, 16F, 17F, 19A, 19F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B and 35F. 207. The immunogenic composition of paragraph 206, which is a 24-valent Streptococcus pneumoniae conjugate composition. 208. The immunogenic composition of any one of paragraphs 129 to 150, further comprising a carbohydrate conjugate from serotypes 2, 3, 7C, 9N, 10B, 15A, 16F, 17F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B and 35F. 209. The immunogenic composition of paragraph 208, which is a 23-valent Streptococcus pneumoniae conjugate composition. 210. The immunogenic composition of any one of paragraphs 129 to 150, further comprising a carbohydrate conjugate from serotypes 2, 3, 7C, 9N, 10B, 15A, 16F, 17F, 19A, 19F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B and 35F. 211. The immunogenic composition of paragraph 210, which is a 23-valent pneumococcal conjugate composition. 212. The immunogenic composition of any one of paragraphs 128 to 211, further comprising at least one, two or three adjuvants. 213. The immunogenic composition of any one of paragraphs 128 to 211, further comprising an adjuvant. 214. The immunogenic composition of any of paragraphs 128 to 211, further comprising two adjuvants. 215. The immunogenic composition of any of paragraphs 128 to 211, further comprising an aluminum salt (aluminum) as an adjuvant. 216. The immunogenic composition of any of paragraphs 128 to 211, further comprising aluminum phosphate as an adjuvant. 217. The immunogenic composition of any of paragraphs 128 to 211, further comprising a saponin-based adjuvant. 218. The immunogenic composition of any of paragraphs 128 to 211, further comprising a QS21-based adjuvant. 219. The immunogenic composition of any of paragraphs 128 to 211, further comprising a CpG oligonucleotide as an adjuvant. 220. The immunogenic composition of any of paragraphs 128 to 219 for use as a vaccine. 221. The immunogenic composition of any of paragraphs 128 to 219 for use in a method of preventing, treating or ameliorating an infection, disease or condition associated with S. pneumoniae serotype 38 in a subject. 222. The immunogenic composition of any of paragraphs 128 to 219 for use in a method of inducing an immune response against S. pneumoniae serotype 38 in a subject, the method comprising administering to the subject an immunologically effective amount of the immunogenic composition. 223. The immunogenic composition of any of paragraphs 128 to 219 for use in a method of preventing infection by S. pneumoniae serotype 38 in a subject, the method comprising administering to the subject an immunologically effective amount of the immunogenic composition. 224. A method for detecting the presence of D-2-acetamido-2,6-dideoxy-xylose-hex-4-ketose (D-Sug) residues in isolated Streptococcus pneumoniae serotype 38 polysaccharides, the method comprising the steps of: a) isolating Streptococcus pneumoniae serotype 38 polysaccharides and b) detecting the presence of D-2-acetamido-2,6-dideoxy-xylose-hex-4-ketose residues in the polysaccharides. 225. The method of paragraph 224, wherein the presence of the D-Sug residue is detected by NMR or mass spectrometry (MS). 226. A method for detecting the presence of N-acetyl-D-fucosamine (D-FucNAc) residues in reduced serotype 38 polysaccharides, the method comprising the steps of: a) reacting isolated S. pneumoniae serotype 38 polysaccharides with a reducing agent, and b) detecting the presence of N-acetyl-D-fucosamine (D-FucNAc) residues in the reduced polysaccharides. 226. The method of paragraph 225, wherein the presence of N-acetyl-D-fucosamine (D-FucNAc) residues is detected by NMR. 227. The method of any of paragraphs 225 to 226, wherein the reducing agent is sodium borohydride ( NaBH4 ). 228. The method of any one of 225 to 227, wherein the isolated S. pneumoniae serotype 38 polysaccharide has been previously treated with an oxidizing agent. 229. A method for detecting the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues in reduced serotype 38 polysaccharides, the method comprising the steps of: a) reacting the isolated S. pneumoniae serotype 38 polysaccharide with a reducing agent, and b) detecting the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues in the reduced polysaccharide. 230. The method of paragraph 229, wherein the presence of the N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue is detected by NMR. 231. The method of paragraph 229, wherein the presence of the N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue is detected by mass spectrometry (MS). 232. The method of any one of paragraphs 229 to 231, wherein the reducing agent is sodium borohydride (NaBH 4 ). 233. The method of any one of paragraphs 229 to 232, wherein the isolated S. pneumoniae serotype 38 polysaccharide has been previously treated with an oxidizing agent. 234. A method for detecting the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfuramine (D-QuiNAc) residues in reduced serotype 38 polysaccharides, the method comprising the steps of: a) reacting isolated Streptococcus pneumoniae serotype 38 polysaccharides with a reducing agent, and b) detecting the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfuramine (D-QuiNAc) residues in the reduced polysaccharide. 235. The method of paragraph 234, wherein the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by NMR. 236. The method of paragraph 234, wherein the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by mass spectrometry (MS). 237. The method of any one of paragraphs 234 to 236, wherein the reducing agent is sodium borohydride (NaBH 4 ). 238. The method of any one of items 234 to 237, wherein the isolated S. pneumoniae serotype 38 polysaccharide has been previously treated with an oxidizing agent. 239. A method for detecting the presence of N-acetyl-D-fucosamine (D-FucNAc) and/or N-acetyl-D-quinolfuramine (D-QuiNAc) residues in a S. pneumoniae serotype 38 glycoconjugate, the method comprising the steps of: a) preparing a S. pneumoniae serotype 38 glycoconjugate, and b) detecting the presence of N-acetyl-D-fucosamine (D-FucNAc) and/or N-acetyl-D-quinolfuramine (D-QuiNAc) residues in the glycoconjugate. 240. The method of paragraph 239, wherein the presence of N-acetyl-D-fucosamine (D-FucNAc) and/or N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by NMR. 241. The method of paragraph 239, wherein the presence of N-acetyl-D-fucosamine (D-FucNAc) and/or N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by mass spectrometry (MS). 242. A method for detecting the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfuramine (D-QuiNAc) residues in a glycoconjugate of Streptococcus pneumoniae serotype 38, the method comprising the steps of: a) preparing a glycoconjugate of Streptococcus pneumoniae serotype 38, and b) detecting the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfuramine (D-QuiNAc) residues in the glycoconjugate. 243. The method of paragraph 242, wherein the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by NMR. 244. The method of paragraph 242, wherein the presence of N-acetyl-D-fucosamine (D-FucNAc) and N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by mass spectrometry (MS). 245. A method for detecting the presence of an N-acetyl-D-fucosamine (D-FucNAc) residue in a glycoconjugate of Streptococcus pneumoniae serotype 38, the method comprising the steps of: a) preparing a glycoconjugate of Streptococcus pneumoniae serotype 38, and b) detecting the presence of an N-acetyl-D-fucosamine (D-FucNAc) residue in the glycoconjugate. 246. The method of paragraph 245, wherein the presence of an N-acetyl-D-fucosamine (D-FucNAc) residue is detected by NMR. 247. The method of paragraph 245, wherein the presence of an N-acetyl-D-fucosamine (D-FucNAc) residue is detected by mass spectrometry (MS). 248. A method for detecting the presence of an N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue in a glycoconjugate of Streptococcus pneumoniae serotype 38, the method comprising the steps of: a) preparing a glycoconjugate of Streptococcus pneumoniae serotype 38, and b) detecting the presence of an N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue in the glycoconjugate. 249. The method of paragraph 248, wherein the presence of an N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residue is detected by NMR. 250. The method of paragraph 248, wherein the presence of N-acetyl-D-quinolfurfurylamine (D-QuiNAc) residues is detected by mass spectrometry (MS). 251. The carbohydrate conjugate of any of paragraphs 43 to 127, for use as an antigen. 252. The carbohydrate conjugate of any of paragraphs 43 to 127, for use as a medicament. 253. The carbohydrate conjugate of any of paragraphs 43 to 127, for use as a vaccine. 254. The carbohydrate conjugate of any of paragraphs 43 to 127, for use in generating an immune response in an individual. 255. The carbohydrate conjugate used in paragraph 254, wherein the individual is a human. 256. The immunogenic composition of any of paragraphs 128 to 219, for use as a medicament. 257. The immunogenic composition of any one of paragraphs 128 to 219 for use as a vaccine. 258. The immunogenic composition of any one of paragraphs 257 for use, wherein the individual is a human. 259. The immunogenic composition of any one of paragraphs 128 to 219 for use in a method of preventing, treating or ameliorating a bacterial infection, disease or condition in an individual. 260. The immunogenic composition of any one of paragraphs 128 to 219 for use in a method of preventing, treating or ameliorating a Streptococcus pneumoniae serotype 38 infection, disease or condition in an individual. 261. The immunogenic composition of any one of paragraphs 128 to 219 for use in a method of inducing an immune response against Streptococcus pneumoniae serotype 38 in an individual. 262. An immunogenic composition as claimed in any one of paragraphs 128 to 219 for use in a method of preventing an individual from being infected by S. pneumoniae serotype 38. 263. An immunogenic composition as claimed in any one of paragraphs 128 to 219 for use in a method of protecting a human susceptible to infection by S. pneumoniae serotype 38. 264. A method of preparing a S. pneumoniae serotype 38 carbohydrate conjugate using click chemistry, the method comprising the steps of: (a) reacting an isolated serotype 38 carbohydrate with a carbonate derivative and an azido linker in an aprotic solvent to produce an activated azido sugar (sugar activation), (b) reacting a carrier protein with a protein having an N-hydroxysuccinimide (NHS) moiety and an alkynyl reagent, wherein the NHS moiety reacts with the amine group to form an amide bond, thereby obtaining an alkynyl-functionalized carrier protein (activation of the carrier protein), and (c) reacting the activated azido sugar of step (a) with the activated alkynyl-carrier protein of step (b) by a Cu+1-mediated azido-alkyne cycloaddition reaction to form a glycoconjugate. 265. The method of paragraph 264, wherein the sugar has been sized to a target molecular weight (MW) range prior to activation (a). 266. The method of paragraph 264, wherein the isolated serotype 38 sugar has been sized prior to activation with a carbonate derivative and an azido linker. 267. The method of paragraph 266, wherein the size of the separated serotype 38 capsular polysaccharide is set to a weight average molecular weight between 50 kDa and 500 kDa. 268. The method of paragraph 266, wherein the size of the separated serotype 38 capsular polysaccharide is set to a weight average molecular weight between 50 kDa and 400 kDa. 268. The method of paragraph 266, wherein the size of the separated serotype 38 capsular polysaccharide is set to a weight average molecular weight between 100 kDa and 500 kDa. 270. The method of paragraph 266, wherein the size of the separated serotype 38 capsular polysaccharide is set to a weight average molecular weight between about 100 kDa and about 400 kDa. 271. The method of paragraph 266, wherein the isolated serotype 38 capsular polysaccharide is sized to have a weight average molecular weight between about 150 kDa and about 300 kDa. 272. The method of paragraph 264, wherein the isolated serotype 38 capsular polysaccharide is not sized prior to activation with a carbonate derivative and an azido linker. 273. The method of any of paragraphs 264 to 272, wherein the carbonate derivative is selected from the group consisting of 1,1'-carbonyldiimidazole (CDI), 1,1'-carbonyl-bis-(1,2,4-triazole) (CDT), N,N'-disuccinimidyl carbonate (DSC), and N-hydroxysuccinimidyl chloroformate. 274. The method of any one of paragraphs 264 to 272, wherein the carbonic acid derivative is 1,1'-carbonyldiimidazole (CDI). 275. The method of any one of paragraphs 264 to 272, wherein the carbonic acid derivative is 1,1'-carbonyl-di-(1,2,4-triazole) (CDT). 276. The method of any one of paragraphs 264 to 272, wherein the carbonic acid derivative is N,N'-disuccinimidyl carbonate (DSC). 277. The method of any one of paragraphs 264 to 272, wherein the carbonic acid derivative is N-hydroxysuccinimidyl chloroformate. 278. The method of any one of paragraphs 264 to 277, wherein the azido linker is a compound of formula (VI), wherein X is selected from the group consisting of CH2 ( CH2 ) n , ( CH2CH2O ) mCH2CH2 , NHCO ( CH2 ) n , NHCO( CH2CH2O ) mCH2CH2 , OCH2 ( CH2 ) n , and O( CH2CH2O ) mCH2CH2 ; wherein n is selected from 1 to 10 and m is selected from 1 to 4. 279. The method of any one of paragraphs 264 to 277, wherein the azido linker is a compound of formula (VI), wherein X is CH2 ( CH2 ) n , and n is selected from 1 to 10. 280. The method of any of paragraphs 264 to 277, wherein the azido linker is a compound of formula (VI), wherein X is (CH 2 CH 2 O) m CH 2 CH 2 , wherein m is selected from 1 to 4. 281. The method of any of paragraphs 264 to 277, wherein the azido linker is a compound of formula (VI), wherein X is NHCO(CH 2 ) n , and n is selected from 1 to 10. 282. The method of any of paragraphs 264 to 277, wherein the azido linker is a compound of formula (VI), wherein X is NHCO(CH 2 CH 2 O) m CH 2 CH 2 , wherein m is selected from 1 to 4. 283. The method of any one of paragraphs 264 to 277, wherein the azido linker is a compound of formula (VI), wherein X is OCH2 ( CH2 ) n , and n is selected from 1 to 10. 284. The method of any one of paragraphs 264 to 277, wherein the azido linker is a compound of formula (VI), wherein X is O( CH2CH2O ) mCH2CH2 , wherein m is selected from 1 to 4. 285. The method of any one of paragraphs 264 to 277 , wherein the azido linker is a compound of formula ( VII ), . 286. The method of any one of paragraphs 264 to 277, wherein the azido linker is 3-azido-propylamine. 287. The method of any one of paragraphs 264 to 286, wherein the reagent having an N-hydroxysuccinimide (NHS) moiety and an alkynyl group is a reagent having an N-hydroxysuccinimide (NHS) moiety and a terminal alkynyl group. 288. The method of any one of paragraphs 264 to 286, wherein the reagent having an N-hydroxysuccinimide (NHS) moiety and an alkynyl group is a reagent having an N-hydroxysuccinimide (NHS) moiety and a cycloalkynyl group. 289. The method of any one of paragraphs 264 to 286, wherein the reagent having an N-hydroxysuccinimide (NHS) moiety and an alkynyl group is a compound of formula (VIII), wherein X is selected from the group consisting of CH2O ( CH2 ) nCH2C =O and CH2O ( CH2CH2O ) m ( CH2 ) nCH2C =O, wherein n is selected from 0 to 10 and m is selected from 0 to 4. 290. The method of any one of paragraphs 264 to 286, wherein the reagent having an N-hydroxysuccinimide (NHS) moiety and an alkynyl group is a compound of formula (VIII), wherein X is CH2O(CH2)nCH2C = O , wherein n is selected from 0 to 10. 291. The method of any one of paragraphs 264 to 286, wherein the reagent having an N-hydroxysuccinimide (NHS) moiety and an alkynyl group is a compound of formula (VIII), wherein X is CH2O(CH2CH2O)m(CH2)nCH2C=O, wherein n is selected from 0 to 10 and m is selected from 0 to 4. 292. The method of any one of paragraphs 264 to 286, wherein the reagent having an N-hydroxysuccinimide (NHS) moiety and an alkynyl group is a compound of formula (IX): . 293. The method of any of paragraphs 264 to 292, wherein step a) comprises reacting a sugar with a carbonic acid derivative, followed by reacting the sugar activated by the carbonic acid derivative with an azido linker in an aprotic solvent to produce an activated azido sugar. 294. The method of any of paragraphs 264 to 293, wherein step a) comprises reacting a sugar with a carbonic acid derivative in an amount between 0.01 and 10 molar equivalents relative to the amount of sugar present in the reaction mixture. 295. The method of any of paragraphs 264 to 294, wherein at step a), the separated sugar is reacted with a carbonic acid derivative in an aprotic solvent. 296. The method of any of paragraphs 264 to 295, wherein the separated sugar is reacted with CDI in an aprotic solvent comprising 0.1% to 1% (v/v) water. 297. The method of any of paragraphs 264 to 296, wherein step a) further comprises reacting the carbonate-activated sugar with an azido linker in an amount between 0.01 and 10 molar equivalents of polysaccharide repeating units (molar equivalents of RU) of the activated sugar. 298. The method of any of paragraphs 264 to 297, wherein the degree of activation of the activated sugar after step a) is between 1.0 and 100%. 299. The method of any of paragraphs 264 to 297, wherein the degree of activation of the activated carbohydrate after step a) is between 5 and 70%. 300. The method of any of paragraphs 264 to 297, wherein the degree of activation of the activated carbohydrate after step a) is between 15 and 50%. 301. The method of any of paragraphs 264 to 297, wherein the degree of activation of the activated carbohydrate after step a) is between 10 and 40%. 302. The method of any of paragraphs 264 to 297, wherein the degree of activation of the activated carbohydrate after step a) is between 5 and 15%. 303. The method of any of paragraphs 264 to 297, wherein the degree of activation of the activated carbohydrate after step a) is between 15 and 35%. 304. The method of any of paragraphs 264 to 297, wherein the degree of activation of the activated sugar after step a) is between 15 and 25%. 305. The method of any of paragraphs 264 to 297, wherein the degree of activation of the activated sugar after step a) is about 25%. 306. The method of any of paragraphs 264 to 305, wherein step b) comprises reacting the carrier protein with a reagent having an N-hydroxysuccinimide (NHS) moiety and an alkynyl group in an amount of 0.1 to 10 molar equivalents relative to the lysine on the carrier. 307. The method of any of paragraphs 264 to 306, wherein the degree of activation of the activated carrier after step b) is between 1 and 50. 308. The method of any one of paragraphs 264 to 306, wherein the degree of activation of the activated support after step b) may be between 5 and 50. 309. The method of any one of paragraphs 264 to 306, wherein the degree of activation of the activated support after step b) is between 10 and 40. 310. The method of any one of paragraphs 264 to 306, wherein the degree of activation of the activated support after step b) is between 15 and 25. 311. The method of any one of paragraphs 264 to 310, wherein the binding reaction c) is carried out in an aqueous buffer. 312. The method of any one of paragraphs 264 to 310, wherein the binding reaction c) is carried out in an aqueous buffer in the presence of copper (I) as a catalyst. 313. The method of any one of paragraphs 264 to 310, wherein the conjugation reaction c) is carried out in an aqueous buffer in the presence of an oxidant and copper (I) as a catalyst. 314. The method of any one of paragraphs 264 to 313, wherein the initial input ratio (weight/weight) of activated azido sugar to activated alkyne-support at step c) is between 0.8 and 1.2. 315. The method of any one of paragraphs 264 to 314, wherein after step c), the unreacted azido groups in the conjugate are capped using a suitable azido capping agent. 316. The method of any of paragraphs 264 to 315, wherein after step c), unreacted alkynyl groups are capped using a suitable alkynyl capping agent. 317. The method of any of paragraphs 283 to 336, wherein the Streptococcus pneumoniae serotype 38 glycoconjugate is as described in any of paragraphs 1 to 57. 318. A method for preparing a Streptococcus pneumoniae serotype 38 glycoconjugate using reductive amination, the method comprising the steps of: (1) oxidizing (activating) serotype 38 purified saccharides, (2) reducing the activated saccharides and a carrier protein to form a saccharide conjugate. 319. The method of paragraph 318, wherein the isolated serotype 38 polysaccharide is sized prior to oxidation. 320. A method for preparing a serotype 38 saccharide conjugate of Streptococcus pneumoniae, comprising the steps of: (a) reacting the serotype 38 saccharide with an oxidizing agent; (b) compounding the activated saccharide of step (a) with a carrier protein; and (c) reacting the compounded activated saccharide and carrier protein with a reducing agent to form a saccharide conjugate. 321. The method of paragraph 320, wherein the oxidizing agent is a periodate. 322. The method of paragraph 321, wherein the degree of oxidation of the activated serotype 38 saccharide is between 2 and 30. 323. The method of paragraph 321, wherein the degree of oxidation (DO) of the activated serotype 38 saccharide is between 10 and 25. 324. The method of paragraph 341, wherein the degree of oxidation (DO) of the activated serotype 38 sugars is between 10 and 25, and wherein the initial input ratio (weight/weight) of the activated serotype 38 sugars to the carrier protein at step b) is between 1.5:1 and 0.5:1. 325. The method of paragraph 321, wherein the degree of oxidation (DO) of the activated serotype 38 sugars is between 10 and 25, and wherein the initial input ratio (weight/weight) of the activated serotype 38 sugars to the carrier protein at step b) is between 1.5:1 and 0.5:1, and wherein the reduction reaction (c) is performed in an aprotic solvent. 326. The method of paragraph 321, wherein the degree of oxidation (DO) of the activated serotype 38 sugar is between 10 and 25, and wherein the initial input ratio (weight/weight) of the activated serotype 38 sugar to the carrier protein at step b) is between 1.5:1 and 0.5:1, and wherein the reduction reaction (c) is performed in a solution consisting essentially of dimethyl sulfoxide (DMSO). 327. The method of paragraph 321, wherein the degree of oxidation (DO) of the activated serotype 38 sugar is between 10 and 25, and wherein the initial input ratio (weight/weight) of the activated serotype 38 sugar to the carrier protein at step b) is between 1.5:1 and 0.5:1, and wherein the reduction reaction (c) is performed in a DMSO (dimethyl sulfoxide) solvent. 328. The method of paragraph 321, wherein the degree of oxidation (DO) of the activated serotype 38 saccharide is between 10 and 25, and wherein the initial input ratio (weight/weight) of the activated serotype 38 saccharide to the carrier protein at step b) is between 1.5:1 and 0.5:1, wherein the reduction reaction (c) is carried out in DMSO (dimethyl sulfoxide) solvent and wherein the reducing agent is sodium cyanoborohydride. 329. The method of any one of paragraphs 320 to 328, wherein the pneumococcal serotype 38 saccharide conjugate is any one of paragraphs 44 to 74. 330. The method of paragraph 328, wherein the pneumococcal serotype 38 saccharide conjugate is any one of paragraphs 44 to 74.

如本文所使用，術語「約」意謂在值之統計學上有意義的範圍內，諸如所陳述之濃度範圍、時間範圍、分子量、溫度或pH。此類範圍可在給定值或範圍之一數量級內，通常在20%內，更通常在10%內，且甚至更通常在5%內或在1%內。有時，此類範圍可能在用於量測及/或測定給定值或範圍之標準方法的典型實驗誤差內。術語「約」所涵蓋之容許偏差將取決於所研究之特定系統，且可由一般熟習此項技術者容易地理解。每當在本申請案中敍述一個範圍時，亦考慮該範圍內之每個數目作為本揭露之一實施例。As used herein, the term "about" means within a statistically significant range of values, such as a stated concentration range, time range, molecular weight, temperature, or pH. Such ranges may be within an order of magnitude of a given value or range, typically within 20%, more typically within 10%, and even more typically within 5% or within 1%. Sometimes, such ranges may be within the typical experimental error of a standard method for measuring and/or determining a given value or range. The allowable deviations covered by the term "about" will depend on the specific system being studied and can be easily understood by those of ordinary skill in the art. Whenever a range is described in this application, each number within the range is also considered as an embodiment of the present disclosure.

本發明人預期，在各情況下，本文中之術語「包含(comprising)」、「包含(comprise)」及「包含(comprises)」視情況可分別替換為術語「基本上由…組成(consisting essentially of)」、「基本上由…組成(consist essentially of)」、「基本上由…組成(consists essentially of)」、「由…組成(consisting of)」、「由…組成(consist of)」及「由…組成(consists of)」。The inventors intend that the terms “comprising,” “comprise,” and “comprises” herein may be replaced with the terms “consisting essentially of,” “consist essentially of,” “consists essentially of,” “consisting of,” “consist of,” and “consists of,” respectively, as appropriate, in each instance.

「免疫原性量」、「免疫有效量」、「治療有效量」、「預防有效量」或「劑量」中之各者在本文中可互換使用，通常係指藉由熟習此項技術者已知之標準分析所量測，足以誘發免疫反應，亦即細胞(T細胞)或體液(B細胞或抗體)反應或兩種反應的抗原或免疫原組合物之量。"Immunogenic amount", "immunologically effective amount", "therapeutically effective amount", "prophylactically effective amount" or "dose" are used interchangeably herein and generally refer to the amount of an antigen or immunogenic composition sufficient to induce an immune response, i.e., a cellular (T cell) or humoral (B cell or antibody) response, or both, as measured by standard assays known to those skilled in the art.

考慮本文件之範圍中之任一者內之任何全數整數作為本揭露之一實施例。Any whole number within any of the ranges of this document is considered as an embodiment of the present disclosure.

在本專利說明書內引用之所有參考文獻或專利申請案均以引用的方式併入本文中。All references or patent applications cited in this patent specification are incorporated herein by reference.

在隨附實例中說明本發明。除非另外詳細描述，否則使用熟習此項技術者所熟知且常用的標準技術進行以下實例。實例為說明性的，而非限制本發明。The present invention is illustrated in the accompanying examples. Unless otherwise described in detail, the following examples are performed using standard techniques that are well known and commonly used by those skilled in the art. The examples are illustrative and not limiting of the present invention.

實例 實例 1 ：血清型 38 莢膜醣，樣品製備 肺炎鏈球菌血清型 38 多醣： 稱取25 mg凍乾天然肺炎鏈球菌血清型38且溶解於1000 µL之D 2O中。在50℃下對樣品進行水浴音波處理20分鐘。使用音波處理2分鐘，以20%振幅功率持續15秒開啟及15秒關閉的循環來對多醣溶液進行尺寸設定。天然樣品及經尺寸設定之樣品的最終濃度為25 mg/mL。將樣品轉移至NMR管中進行NMR資料收集及分析。 Examples Example 1 : Serotype 38 Capsular Saccharide, Sample Preparation Streptococcus pneumoniae serotype 38 polysaccharide: Weigh 25 mg of freeze-dried native Streptococcus pneumoniae serotype 38 and dissolve in 1000 µL of D 2 O. Sonicate the sample in a water bath at 50°C for 20 minutes. Size the polysaccharide solution using sonication for 2 minutes with a cycle of 15 seconds on and 15 seconds off at 20% amplitude power. The final concentration of the native and sized samples was 25 mg/mL. Transfer the sample to an NMR tube for NMR data collection and analysis.

去乙醯化肺炎鏈球菌血清型 38 多醣 ：在室溫下用0.25N NH 4OH在恆定攪拌下培育24小時後，25 mg天然血清型38多醣完全去乙醯化。接著，使樣品用水透析且凍乾。接著將凍乾多醣再懸浮於1000 µL之D 2O中。將樣品轉移至NMR管中進行NMR資料收集及分析。 Deacetylated S. pneumoniae serotype 38 polysaccharide : 25 mg of native serotype 38 polysaccharide was completely deacetylated after incubation with 0.25N NH 4 OH at room temperature with constant stirring for 24 hours. The sample was then dialyzed against water and lyophilized. The lyophilized polysaccharide was then resuspended in 1000 µL of D 2 O. The sample was transferred to an NMR tube for NMR data collection and analysis.

經還原之肺炎鏈球菌血清型 38 多醣 ：在室溫下用1莫耳當量之NaBH 4在恆定攪拌下培育18小時後，25 mg天然血清型38多醣完全還原。接著，使樣品用水透析且凍乾。接著將凍乾多醣再懸浮於1000 µL之D 2O中。將樣品轉移至NMR管中進行NMR資料收集及分析。 Reduced S. pneumoniae serotype 38 polysaccharide : 25 mg of native serotype 38 polysaccharide was completely reduced after incubation with 1 mol equivalent of NaBH 4 at room temperature under constant stirring for 18 hours. The sample was then dialyzed against water and freeze-dried. The freeze-dried polysaccharide was then resuspended in 1000 µL of D 2 O. The sample was transferred to an NMR tube for NMR data collection and analysis.

實例 2 ： 藉由 NMR 實驗對肺炎鏈球菌血清型 38 多醣進行結構闡明進行一維(1D)及二維(2D) NMR實驗。收集1D 1H光譜以鑑別分子中各質子之化學位移指紋。收集2D同核及異核光譜以鑑別分子中之質子與碳之間的化學鍵模式，繪製血清型38聚合物重複單元中代表性糖之主鏈、糖間糖苷鍵，且鑑別糖主鏈上不同官能基之位置。 Example 2 : Structural Elucidation of S. pneumoniae Serotype 38 Polysaccharide by NMR Experiments One-dimensional (1D) and two-dimensional (2D) NMR experiments were performed. 1D 1 H spectra were collected to identify the chemical shift fingerprints of each proton in the molecule. 2D homonuclear and heteronuclear spectra were collected to identify the chemical bonding pattern between protons and carbons in the molecule, to map the main chain of representative sugars in the repeating unit of the serotype 38 polymer, the glycosidic bonds between sugars, and to identify the positions of different functional groups on the sugar main chain.

在75℃下使用配備有BBO低溫探針之Bruker 600 MHz光譜儀收集所有1D及2D資料。NMR資料使用NvX處理且使用MestraNova及NMRViewJ (NvJ)軟體分析( Methods Mol . Biol. 278, 313, 2004)。使用設定為0 ppm之TMS信號來參考質子信號，而碳信號間接地參考TMS信號(0 ppm)。 All 1D and 2D data were collected at 75°C using a Bruker 600 MHz spectrometer equipped with a BBO cryoprobe. NMR data were processed using NvX and analyzed using MestraNova and NMRViewJ (NvJ) software ( Methods Mol . Biol . 278, 313, 2004). Proton signals were referenced to the TMS signal set to 0 ppm, while carbon signals were indirectly referenced to the TMS signal (0 ppm).

用32次掃描收集1D質子資料，其中再循環延遲為10s。記錄以下2D NMR實驗： 1H- 1H同核COSY (相關光譜法，2048×512：分別在 1H及 13C維度上的點總數，2次掃描，再循環延遲1s)， 1H- 13C HSQC (異核單量子相干，2048×256：分別在 1H及 13C維度上的點總數，4次掃描，再循環延遲1s)， 1H- 13C HSQC-TOCSY (異核單量子相干-總相關光譜法，2048×256：分別在 1H及 13C維度上的點總數，48次掃描，再循環延遲1s，混合時間為120ms)， 1H- 13C HMBC (異核多鍵相關，2048×256：分別在 1H及 13C維度上的點總數，長程J約5-10 Hz，48次掃描，再循環延遲1s)及 1H- 13C HSQC-COSY (異核單量子相干-相關光譜法，2048×256：分別在 1H及 13C維度上的點總數，48次掃描，再循環延遲1s)， 1H- 13C CLIP-HSQC ( 1H去耦異核單量子相干，2048×256：分別在 1H及 13C維度上的點總數，8次掃描，再循環延遲1s)。用256次掃描收集1D 31P資料，其中再循環延遲為5s。 1H- 31P HMBC (異核多鍵相關，2048×32：分別在 1H及 31P維度上的點總數，長程J約5-10 Hz)。 1D proton data were collected using 32 scans with a recycle delay of 10 s. The following 2D NMR experiments were recorded: 1 H- 1 H homonuclear COSY (correlation spectroscopy, 2048 × 512: total number of points in 1 H and 13 C dimensions, 2 scans, recycle delay 1 s), 1 H- 13 C HSQC (heteronuclear single quantum coherence, 2048 × 256: total number of points in 1 H and 13 C dimensions, 4 scans, recycle delay 1 s), 1 H- 13 C HSQC-TOCSY (heteronuclear single quantum coherence-total correlation spectroscopy, 2048 × 256: total number of points in 1 H and 13 C dimensions, 48 scans, recycle delay 1 s, mixing time 120 ms), 1 H- 13 C HMBC (heteronuclear multibond correlation, 2048×256: total number of points in 1 H and 13 C dimensions, respectively, long range J about 5-10 Hz, 48 scans, recycle delay 1s) and 1H- 13C HSQC-COSY (heteronuclear single quantum coherence-correlation spectroscopy, 2048×256: total number of points in 1 H and 13 C dimensions, respectively, 48 scans, recycle delay 1s), 1H- 13C CLIP-HSQC ( 1H decoupled heteronuclear single quantum coherence, 2048×256: total number of points in 1 H and 13 C dimensions, respectively, 8 scans, recycle delay 1s). 1D 31 P data were collected with 256 scans with a recycle delay of 5 s. 1 H- 31 P HMBC (heteronuclear multibond correlation, 2048 × 32: total number of points in the 1 H and 31 P dimensions, respectively, long range J about 5-10 Hz).

已發現兩種結構取決於起始菌株(兩種形式：命名為絲胺酸形式及甘胺酸形式)。肺炎鏈球菌血清型38絲胺酸形式及甘胺酸形式之化學結構分別展示於圖1A及圖1B中。血清型38多醣為五醣：α-D-葡糖胺(A)、α-D-2-乙醯胺基-2,6-二去氧基-木糖-己-4-酮糖(α-D-Sug) (B)、O-乙醯化β-D-半乳糖(C)、α-D-半乳糖(D)及β-D-呋喃半乳糖(E)。殘基C (O-乙醯化β-D-半乳糖)在位置6處進一步與絲胺酸或甘胺酸胺基酸連接，由此表示多醣之血清型38絲胺酸及甘胺酸形式。Two structures have been found depending on the starting strain (two forms: named serine form and glycine form). The chemical structures of the serine form and glycine form of Streptococcus pneumoniae serotype 38 are shown in Figure 1A and Figure 1B, respectively. The serotype 38 polysaccharide is a pentasaccharide: α-D-glucosamine (A), α-D-2-acetamido-2,6-dideoxy-xylose-hex-4-ketose (α-D-Sug) (B), O-acetylated β-D-galactose (C), α-D-galactose (D) and β-D-furanose (E). The residual C (O-acetylated β-D-galactose) is further linked to the serine or glycine amino acid at position 6, thereby representing the serotype 38 serine and glycine forms of the polysaccharide.

血清型38多醣之β-D-Gal p4OAc,6Ser (殘基C)在碳位置4處經O-乙醯化，且乙醯化水平為約94%。 The β-D-Gal p 4OAc,6Ser (residue C) of serotype 38 polysaccharide is O-acetylated at carbon position 4, and the acetylation level is about 94%.

對血清型 38 多醣之 2D NMR 分析藉由對重複單元中存在的各糖單元的所有共振進行完整分配來達成經尺寸設定之血清型38 (絲胺酸形式及甘胺酸形式)多醣的完整結構闡明。使用2D異核NMR實驗分配糖單元中之各者，該等實驗使用短程 1H- 1H (HSQC-COSY)、長程(多鍵) 1H- 1H (HSQC-TOCSY)或 13C- 13C (HMBC)相關性，來繪製糖環內的共振。 2D NMR analysis of serotype 38 polysaccharides Complete structural elucidation of the sized serotype 38 (serine form and glycine form) polysaccharide was achieved by complete assignment of all resonances for each sugar unit present in the repeat unit. Each of the sugar units was assigned using 2D heteronuclear NMR experiments that mapped resonances within the sugar ring using short-range 1 H- 1 H (HSQC-COSY), long-range (multi-bond) 1 H- 1 H (HSQC-TOCSY), or 13 C- 13 C (HMBC) correlations.

2D HSQC實驗提供直接鍵結之 1H與 13C信號之間的相關性且對於分析多醣而言極其靈敏。2D HSQC-COSY產生 1H- 1H短程且HSQC-TOCSY產生 1H- 1H長程(混合時間約120 ms)質子相關性且極適用於在糖環內提供共振。類似地，逆向偵測的2D 1H- 13C HMBC實驗亦在質子與碳原子之間產生交叉峰，該等交叉峰係透過多個碳鍵進行長程標量耦合的。此等交叉峰之強度反映在 2J C , H或 3J C , H值中。通常，自變旋異構碳至位置3及位置5處的碳之間觀察到三個鍵的相關性。透過糖苷鍵與相鄰碳之相關性亦相當重要，此有助於糖單元之連續分配及其連接性。 2D HSQC experiments provide correlations between directly bonded 1 H and 13 C signals and are extremely sensitive for analyzing polysaccharides. 2D HSQC-COSY produces 1 H- 1 H short-range and HSQC-TOCSY produces 1 H- 1 H long-range (mixing time about 120 ms) proton correlations and are very suitable for providing resonances within sugar rings. Similarly, the inversely detected 2D 1 H- 13 C HMBC experiment also produces cross peaks between protons and carbon atoms, which are long-range scalar coupled through multiple carbon bonds. The intensity of these cross peaks is reflected in the 2 J C , H or 3 J C , H values. Typically, three-bond correlations are observed between the autorotatable isomeric carbon to the carbon at position 3 and position 5. The association with adjacent carbons through the glycosidic bond is also important, which helps the sequential distribution of sugar units and their connectivity.

藉由比較來自短程(HSQC-COSY)及長程(HSQC-TOCSY)之共振來分配水合物及酮狀態的所有糖主鏈共振。All sugar backbone resonances were assigned to the hydrate and keto states by comparing the resonances from short range (HSQC-COSY) and long range (HSQC-TOCSY).

針對絲胺酸形式及甘胺酸形式，血清型38多醣之 1H- 13C HSQC相關性譜與血清型38重複單元中所有糖的共振的完整分配列於表1中。 The 1 H- 13 C HSQC correlation spectra of the serotype 38 polysaccharide are listed in Table 1 for both the serine form and the glycine form along with the complete assignment of resonances for all sugars in the serotype 38 repeat unit.

表1.血清型38重複單元之所有糖之化學位移分配(針對絲胺酸形式及甘胺酸形式)： 絲胺酸形式 甘胺酸形式 1H (ppm) 13C (ppm) 1H (ppm) 13C (ppm) D-a-Glc pNAc (A) A1 5.24 99.9 5.27 99.7 (水合物形式) A2 4.11 53.9 4.12 53.9 A3 4.11 77.6 4.05 77.3 A4 3.71 68.6 3.71 68.5 A5 3.71 72.9 3.71 72.9 A6 3.88 60.9 3.89 60.9 ANAc 2.08 22.8 2.07 22.7 CO 174.4 174.4 D-a-Sug (B) B1 4.80 98.7 4.80 98.7 (水合物形式) B2 4.24 51.8 4.24 51.8 B3 3.91 78.7 3.93 78.3 B4 94.8 94.9 B5 4.33 69.6 4.33 69.6 B6 1.19 11.2 1.19 11.2 BNAc 2.06 22.8 2.07 22.7 CO 174.3 174.0 D-b-Gal p4OAc,6Ser (C) C1 4.85 101.5 4.83 101.3 D-b-Gal p4OAc,6Gly (C) C2 3.89 73.6 3.89 73.6 (水合物形式) C3 4.25 74.2 4.23 74.2 C4 5.87 66.8 5.86 66.8 C5 4.40 73.8 4.41 73.8 C4OAc 2.12 20.5 2.12 20.6 CO 172.8 172.9 Ser-Ca 4.27 57.5 Ser-Cb 3.83 62.7 Gly-H1 3.86 43.7 Gly-H2 3.79 43.7 D-a-Gal p(D) D1 5.24 94.6 5.25 94.6 (水合物形式) D2 3.93 68.3 3.91 68.3 D3 3.94 69.3 3.94 69.4 D4 4.01 78.2 4.00 78.1 D5 4.12 72.3 4.11 72.3 D6 3.70 60.8 3.71 60.8 D-b-Gal f(E) G1 5.16 108.3 5.16 108.3 (水合物形式) G2 4.03 83.0 4.02 83.0 G3 4.00 78.1 4.00 78.1 G4 4.06 81.6 4.06 81.7 G5 3.87 70.6 3.87 70.7 G6 3.74 64.1 3.74 64.1 D-a-Glc pNAc (A) A1 酮 5.06 97.3 5.07 97.2 (酮形式) A2 酮 4.15 53.6 4.15 53.5 A3 酮 3.98 78.2 3.99 78.3 A4 酮 3.68 68.7 3.67 68.6 A5 酮 3.67 73.2 3.67 73.2 D-a-Sug (B) B1 酮 5.05 98.6 5.05 98.6 (酮形式) B2 酮 4.48 55.6 4.50 55.6 B3 酮 4.69 77.1 4.69 76.9 B4 酮 204.8 204.8 B5 酮 5.06 70.7 5.08 70.7 B6 酮 1.22 12.7 1.22 12.8 Table 1. Chemical shift assignments of all sugars of serotype 38 repeat units (for serine and glycine forms): Serine form Glycine form 1H (ppm) 13C (ppm) 1H (ppm) 13C (ppm) Da-Glc p NAc (A) A1 5.24 99.9 5.27 99.7 (Hydrate form) A2 4.11 53.9 4.12 53.9 A3 4.11 77.6 4.05 77.3 A4 3.71 68.6 3.71 68.5 A5 3.71 72.9 3.71 72.9 A6 3.88 60.9 3.89 60.9 ANAc 2.08 22.8 2.07 22.7 CO 174.4 174.4 Da-Sug (B) B1 4.80 98.7 4.80 98.7 (Hydrate form) B2 4.24 51.8 4.24 51.8 B3 3.91 78.7 3.93 78.3 B4 94.8 94.9 B5 4.33 69.6 4.33 69.6 B6 1.19 11.2 1.19 11.2 BNAc 2.06 22.8 2.07 22.7 CO 174.3 174.0 Db-Gal p 4OAc,6Ser (C) C1 4.85 101.5 4.83 101.3 Db-Gal p 4OAc,6Gly (C) C2 3.89 73.6 3.89 73.6 (Hydrate form) C3 4.25 74.2 4.23 74.2 C4 5.87 66.8 5.86 66.8 C5 4.40 73.8 4.41 73.8 C4 2.12 20.5 2.12 20.6 CO 172.8 172.9 Ser-Ca 4.27 57.5 Ser-Cb 3.83 62.7 Gly-H1 3.86 43.7 Gly-H2 3.79 43.7 Da-Gal p (D) D1 5.24 94.6 5.25 94.6 (Hydrate form) D2 3.93 68.3 3.91 68.3 D3 3.94 69.3 3.94 69.4 D4 4.01 78.2 4.00 78.1 D5 4.12 72.3 4.11 72.3 D6 3.70 60.8 3.71 60.8 Db-Gal f (E) G1 5.16 108.3 5.16 108.3 (Hydrate form) G2 4.03 83.0 4.02 83.0 G3 4.00 78.1 4.00 78.1 G4 4.06 81.6 4.06 81.7 G5 3.87 70.6 3.87 70.7 G6 3.74 64.1 3.74 64.1 Da-Glc p NAc (A) A1 Ketone 5.06 97.3 5.07 97.2 (Keto form) A2 Ketone 4.15 53.6 4.15 53.5 A3 Ketone 3.98 78.2 3.99 78.3 A4 Ketone 3.68 68.7 3.67 68.6 A5 Ketone 3.67 73.2 3.67 73.2 Da-Sug (B) B1 Ketone 5.05 98.6 5.05 98.6 (Keto form) B2 Ketone 4.48 55.6 4.50 55.6 B3 Ketone 4.69 77.1 4.69 76.9 B4 Ketone 204.8 204.8 B5 Ketone 5.06 70.7 5.08 70.7 B6 Ketone 1.22 12.7 1.22 12.8

實例 3 ： 藉由 NMR 實驗對 O - 乙醯化及去 O - 乙醯化血清型 38 ( 絲胺酸形式 ) 多醣進行結構比較乙醯化(下圖)及去乙醯化(上圖)血清型38多醣之1D 1H及 13C NMR光譜之比較示於圖3及圖4中。所有共振標註於1D光譜中，且一些關鍵共振標註於 13C光譜中。乙酸酯基團之甲基、羰基碳共振及共振β-D-Gal p4OAc,6Ser糖之C4均不存在於去乙醯化血清型38多醣中。 Example 3 : Structural comparison of O - acetylated and de- O - acetylated serotype 38 ( serine form ) polysaccharides by NMR experiments Comparison of 1D 1 H and 13 C NMR spectra of acetylated (lower panel) and de-acetylated (upper panel) serotype 38 polysaccharides is shown in Figures 3 and 4. All resonances are annotated in the 1D spectrum, and some key resonances are annotated in the 13 C spectrum. The methyl and carbonyl carbon resonances of the acetate group and the resonance C4 of β-D-Gal p 4OAc,6Ser sugar are not present in the de-acetylated serotype 38 polysaccharide.

實例 4 ： 藉由 NMR 實驗對經還原之血清型 38 多醣 ( 絲胺酸形式 ) 進行結構表徵可使用過碘酸鈉(NaIO 4)進行多醣之活化以進行結合，該過碘酸鈉在聚合物鏈中之選擇性糖處產生一級醛。此經活化之多醣隨後與載體蛋白結合。結合後的關鍵單元操作之一為使用硼氫化鈉(NaBH 4)還原未結合之活化多醣。然而，已發現在此過程期間，血清型38中存在的酮糖(Sug)將被還原。需要理解用NaBH 4還原之後的血清型38的潛在結構變化及其對醣結合物之潛在影響。 Example 4 : Structural Characterization of Reduced Serotype 38 Polysaccharide ( Serine Form ) by NMR Experiments The polysaccharide can be activated for conjugation using sodium periodate (NaIO 4 ), which generates a primary aldehyde at a selective sugar in the polymer chain. This activated polysaccharide is then conjugated to a carrier protein. One of the key unit operations after conjugation is the reduction of the unconjugated activated polysaccharide using sodium borohydride (NaBH 4 ). However, it has been found that during this process, ketoses (Sug) present in serotype 38 will be reduced. It is necessary to understand the potential structural changes of serotype 38 after reduction with NaBH 4 and its potential impact on glycoconjugates.

圖5顯示經還原之血清型38多醣的1D 1H光譜(按實例1中所揭露獲得)。在用硼氫化鈉(NaBH 4)還原後，酮糖(Sug)被還原以形成：不同比率之Fuc pNAc及Qui pNAc糖。來自Fuc pNAc及Qui pNAc殘基之甲基信號下的面積已用於確定具有Fuc pNAc或Qui pNAc之多醣鏈的群體(使用2D 1H- 13C HSQC光譜)。在經還原之38多醣的2D HSQC光譜中，來自Sug殘基之甲基信號已完全消失，表明多醣完全還原。經還原之血清型38多醣在重複單元中分別顯示68%與32%之Fuc pNAc及Qui pNAc糖的比率。 Figure 5 shows the 1D 1 H spectrum of reduced serotype 38 polysaccharide (obtained as disclosed in Example 1). After reduction with sodium borohydride (NaBH 4 ), ketose (Sug) was reduced to form: Fuc p NAc and Qui p NAc sugars in different ratios. The area under the methyl signals from Fuc p NAc and Qui p NAc residues has been used to determine the population of polysaccharide chains with Fuc p NAc or Qui p NAc (using 2D 1 H- 13 C HSQC spectrum). In the 2D HSQC spectrum of reduced 38 polysaccharide, the methyl signals from the Sug residues have completely disappeared, indicating that the polysaccharide is completely reduced. The reduced serotype 38 polysaccharide showed a ratio of 68% and 32% of Fuc p NAc and Qui p NAc sugars in the repeat unit, respectively.

未經還原及經還原之血清型38多醣的1D 13C光譜的比較清楚地顯示了在用NaBH 4還原時所觀測到的化學共振的差異。 Comparison of the 1D 13 C spectra of the unreduced and reduced serotype 38 polysaccharide clearly shows the differences in the chemical resonances observed upon reduction with NaBH 4 .

經還原之38血清型的結構於圖6示出示。The structure of the reduced serotype 38 is shown in FIG6 .

實例 5 ： 使用點擊化學結合肺炎鏈球菌血清型 38 莢膜多醣1.  血清型38莢膜多醣之機械尺寸設定 使天然多醣在15000 psi壓力下經歷機械尺寸設定以減小MW尺寸。進行尺寸設定研究以鑑定達到140-200 kDa的目標多醣MW所需的通過次數。 Example 5 : Binding of S. pneumoniae serotype 38 capsule polysaccharide using click chemistry 1. Mechanical sizing of serotype 38 capsule polysaccharide The native polysaccharide was subjected to mechanical sizing at 15,000 psi pressure to reduce the MW size. Sizing studies were performed to identify the number of passes required to achieve a target polysaccharide MW of 140-200 kDa.

2.  用疊氮基連接子活化血清型38莢膜多醣 將400 mg血清型38多醣(173 kDa)與1.2g咪唑混合，隨後冷凍且凍乾。凍乾後，將凍乾之多醣用200 mL無水DMSO復原。隨後使反應混合物升溫至35℃，且隨後添加CDI (100 mg/mL於DMSO中，1.25 mL，2 MEq)。在35℃下攪拌反應混合物3小時。在將反應混合物冷卻至23℃之後，添加4 mL (2% v/v)注射用水以淬滅游離CDI，且在23℃下攪拌30分鐘。 2. Activation of serotype 38 capsular polysaccharide with an azido linker 400 mg of serotype 38 polysaccharide (173 kDa) was mixed with 1.2 g of imidazole, then frozen and lyophilized. After lyophilization, the lyophilized polysaccharide was reconstituted with 200 mL of anhydrous DMSO. The reaction mixture was then warmed to 35°C, and CDI (100 mg/mL in DMSO, 1.25 mL, 2 MEq) was then added. The reaction mixture was stirred at 35°C for 3 hours. After the reaction mixture was cooled to 23°C, 4 mL (2% v/v) of water for injection was added to quench free CDI, and stirred at 23°C for 30 minutes.

向反應混合物中添加76 µL之3-疊氮基-1-丙胺(2 MEq)及110 µL之三乙胺(2 MEq)，且隨後在23℃下攪拌20小時。在反應之後，用在生理食鹽水(pH 7.0)中冷卻之10 mM SPB稀釋混合物(5次)。隨後藉由UF/DF使用10K MWCO PES膜相對於含10 mM SPB之生理食鹽水(pH 7.0) (30×體積)純化經稀釋之反應混合物。76 µL of 3-azido-1-propylamine (2 MEq) and 110 µL of triethylamine (2 MEq) were added to the reaction mixture, and then stirred at 23°C for 20 hours. After the reaction, the mixture was diluted with 10 mM SPB cooled in saline (pH 7.0) (5 times). The diluted reaction mixture was then purified by UF/DF using a 10K MWCO PES membrane against 10 mM SPB in saline (pH 7.0) (30× volume).

3.  用炔烴NHS酯將SCP或CRM 197活化為炔烴-SCP及炔烴-CRM 197 SCP 之活化 向SCP溶液(2 g，116 mL)中添加284 mL WFI及100 mL 0.5 M磷酸鈉緩衝液(SPB) (pH 8.3)，獲得4 mg/mL的最終濃度以及0.1 M的離子強度以用於反應。在冷卻至8℃之後，向反應混合物中逐滴添加9.2 mL (相對於SCP上之離胺酸為0.5 MEq)之3-炔丙氧基-丙酸NHS酯(POPS) (20 mg/mL於DMSO中)，將反應溫度維持在8 ± 3℃。將反應混合物在10℃下攪拌30分鐘之後，藉由UF/DF使用10K MWCO PES膜相對於含10 mM SPB之生理食鹽水(pH 7.0) (30×透濾體積)對反應混合物進行純化。獲取418 mL滲餘物且添加62.7 gm蔗糖(15% w/v)，且接著進行0.22 μm (Millipak 60)過濾。使用Lowry檢定法分析滲餘物以進行蛋白質定量，且使用SEC-MALLS分析Mw。 3. Activation of SCP or CRM 197 to Alkyne-SCP and Alkyne-CRM 197 Activation of SCP with Alkyne NHS Ester To a SCP solution (2 g, 116 mL) were added 284 mL of WFI and 100 mL of 0.5 M sodium phosphate buffer (SPB) (pH 8.3) to obtain a final concentration of 4 mg/mL and an ionic strength of 0.1 M for the reaction. After cooling to 8°C, 9.2 mL (0.5 MEq relative to lysine on SCP) of 3-propargyloxy-propionic acid NHS ester (POPS) (20 mg/mL in DMSO) was added dropwise to the reaction mixture, maintaining the reaction temperature at 8 ± 3°C. After stirring the reaction mixture at 10°C for 30 minutes, the reaction mixture was purified by UF/DF using a 10K MWCO PES membrane against 10 mM SPB in saline (pH 7.0) (30× filter volume). 418 mL of the raffinate was obtained and 62.7 gm sucrose (15% w/v) was added and then filtered at 0.22 μm (Millipak 60). The raffinate was analyzed using the Lowry assay for protein quantification and SEC-MALLS for Mw.

CRM 197 之活化 CRM 197之活化類似於SCP之活化，不同之處在於連接子之量相對於CRM 197上之離胺酸經調適為0.5 MEq。 Activation of CRM 197 The activation of CRM 197 was similar to that of SCP, except that the amount of linker relative to lysine on CRM 197 was adjusted to 0.5 MEq.

4.  點擊結合 向血清型38疊氮基多醣(如上述第2點處獲得) (62.5 mL)與炔烴-SCP (如上述第3點處獲得) (31.2 mL)或炔烴-CRM 197之混合物中添加1 M SPB (pH 7) 11 mL及5 mL WFI以製備具有100 mM SPB (pH 7)之離子強度的1-2 g/L反應混合物。在室溫下攪拌混合物1小時，隨後添加硫酸銅(31 µmoL，6.2 mL)及參(3-羥基丙基三唑基甲基)胺(THPTA) (155 µmoL，6.2 mL)、胺基胍(1240 µmoL，12.4 mL)及抗壞血酸鈉(1240 µmoL，12.4 mL)之混合物。在室溫下攪拌反應混合物1.5小時。 4. Click conjugation To a mixture of serotype 38 azidopolysaccharide (obtained as above at point 2) (62.5 mL) and alkynyl-SCP (obtained as above at point 3) (31.2 mL) or alkynyl-CRM 197 , 1 M SPB (pH 7) 11 mL and 5 mL WFI were added to prepare a 1-2 g/L reaction mixture with an ionic strength of 100 mM SPB (pH 7). The mixture was stirred at room temperature for 1 hour, and then copper sulfate (31 µmoL, 6.2 mL) and a mixture of tris(3-hydroxypropyltriazolylmethyl)amine (THPTA) (155 µmoL, 6.2 mL), aminoguanidine (1240 µmoL, 12.4 mL) and sodium ascorbate (1240 µmoL, 12.4 mL) were added. The reaction mixture was stirred at room temperature for 1.5 hours.

在1.5小時之後，將36 µL炔丙醇(每炔烴20 MEq)添加至反應混合物中以封端多醣上的未反應的疊氮化物。隨後添加額外點擊試劑(一半量)以增強封端反應；5 mM硫酸銅3.1 mL與25 mM THPTA 3.1 mL之混合物、100 mM胺基胍(與銅混合物溶液相同的體積)及100 mM抗壞血酸鹽(與銅混合物溶液相同的體積)。攪拌反應混合物一小時。After 1.5 hours, 36 µL of propargyl alcohol (20 MEq per alkyne) was added to the reaction mixture to cap the unreacted azide on the polysaccharide. Additional click reagents (half the amount) were then added to enhance the capping reaction; a mixture of 3.1 mL of 5 mM copper sulfate and 3.1 mL of 25 mM THPTA, 100 mM aminoguanidine (same volume as the copper mixture solution), and 100 mM ascorbate (same volume as the copper mixture solution). The reaction mixture was stirred for one hour.

隨後，添加115 µL之3-疊氮基-丙醇(每炔烴40 MEq)以封端未反應的炔烴SCP (或CRM 197)且再攪拌一小時。藉由UF/DF使用100K MWCO RC膜相對於含10 mM EDTA + 10 mM SPB之生理食鹽水(pH 7.0) (30×透濾體積)，接著藉由第2代UF/DF相對於含5 mM丁二酸鹽之生理食鹽水(pH 6.0) (20×透濾體積)來純化反應混合物。在UF/DF之後，滲餘物通過0.22 µm過濾器(Millipak 40)進行過濾，且隨後進行分析。 Subsequently, 115 µL of 3-azido-propanol (40 MEq per alkyne) was added to cap the unreacted alkyne SCP (or CRM 197 ) and stirred for another hour. The reaction mixture was purified by UF/DF using a 100K MWCO RC membrane versus 10 mM EDTA + 10 mM SPB in saline (pH 7.0) (30× filter volume), followed by second generation UF/DF versus 5 mM succinate in saline (pH 6.0) (20× filter volume). After UF/DF, the raffinate was filtered through a 0.22 µm filter (Millipak 40) and subsequently analyzed.

表2顯示所獲得的一些結合物的屬性(第4行及第5行)。Table 2 shows the properties of some of the conjugates obtained (rows 4 and 5).

實例 6 ： 使用還原胺化化學方法結合肺炎鏈球菌血清型 38 莢膜多醣1.  使用過碘酸鈉氧化血清型38莢膜多醣(經氧化之多醣) 將870 mL經機械尺寸設定之血清型38多醣(173kDa，3.0 g)與555 mL WFI及75 mL 1M磷酸鉀緩衝液(KPB) (pH 6)混合，以製備2 mg/mL濃度及50 mM離子強度溶液。在將pH值調節至6.0之後，將過碘酸鈉(0.1 MEq)添加至反應混合物中。反應混合物在5℃下攪拌20小時，接著藉由UF/DF使用10K MWCO PES膜相對於WFI (20X, v/v)進行純化且分析。 Example 6 : Binding of Streptococcus pneumoniae serotype 38 capsular polysaccharide using reductive amination chemistry 1. Oxidation of serotype 38 capsular polysaccharide using sodium periodate (oxidized polysaccharide) 870 mL of mechanically sized serotype 38 polysaccharide (173 kDa, 3.0 g) was mixed with 555 mL of WFI and 75 mL of 1 M potassium phosphate buffer (KPB) (pH 6) to prepare a 2 mg/mL concentration and 50 mM ionic strength solution. After adjusting the pH to 6.0, sodium periodate (0.1 MEq) was added to the reaction mixture. The reaction mixture was stirred at 5 °C for 20 h, then purified and analyzed by UF/DF using a 10K MWCO PES membrane versus WFI (20X, v/v).

2.  在水性系統中使用還原胺化化學方法(RAC) (RAC-Aq)進行結合 將血清型38經氧化之多醣(參見步驟1) (150 mg，25×蔗糖)與CRM 197300 mg (SPR輸入比率：0.5)混合，且接著凍乾(共凍乾)。 2. Conjugation using reductive amination chemistry (RAC) in an aqueous system (RAC-Aq) Serotype 38 oxidized polysaccharide (see step 1) (150 mg, 25× sucrose) was mixed with CRM 197 300 mg (SPR input ratio: 0.5) and then freeze-dried (co-lyophilized).

將凍乾混合物在30 mL 0.1 M KPB (pH 7.5)中復原。將溶液之pH值調節至6。在將反應溶液加熱至30℃之後，添加氰基硼氫化鈉(100 mg/mL於0.15 M KPB中，pH 7.5，老化3小時) 91 µL (1 MEq)。藉由HPLC監測反應，且在72小時之後，添加另一莫耳當量的氰基硼氫化鈉(100 mg/mL於0.15 M KPB中，pH 7.5，老化3小時) 91 µL (1 MEq)。封端24小時。The lyophilized mixture was reconstituted in 30 mL 0.1 M KPB (pH 7.5). The pH of the solution was adjusted to 6. After the reaction solution was heated to 30 °C, sodium cyanoborohydride (100 mg/mL in 0.15 M KPB, pH 7.5, aged 3 hours) 91 µL (1 MEq) was added. The reaction was monitored by HPLC, and after 72 hours, another molar equivalent of sodium cyanoborohydride (100 mg/mL in 0.15 M KPB, pH 7.5, aged 3 hours) 91 µL (1 MEq) was added. Capping was performed for 24 hours.

隨後用300 mL預冷卻的含5 mM丁二酸鹽之生理食鹽水(pH 6.0)稀釋反應混合物，且藉由UF/DF使用100K MWCO RC膜相對於含5 mM丁二酸鹽之生理食鹽水(pH 6.0) (50×透濾體積)進行純化。The reaction mixture was then diluted with 300 mL of pre-chilled saline (pH 6.0) containing 5 mM succinate and purified by UF/DF using a 100K MWCO RC membrane versus 5 mM succinate in saline (pH 6.0) (50× filter volume).

3.  在DMSO中使用還原胺化化學方法(RAC) (RAC-DMSO)進行結合，僅用NaCNBH 3作為還原劑及封端劑(約30%還原，表2之第2行) 將血清型38經氧化之多醣(參見步驟1) (200 mg)與蔗糖5.0 g混合，且隨後凍乾。將凍乾經活化氧化之多醣接著在室溫下用DMSO 100 mL進行復原。且凍乾的CRM 197(200 mg)亦用DMSO 100 mL進行復原。 3. Reductive amination chemistry (RAC) in DMSO (RAC-DMSO) was used for conjugation, using only NaCNBH 3 as reducing agent and capping agent (about 30% reduction, row 2 of Table 2) Serotype 38 oxidized polysaccharide (see step 1) (200 mg) was mixed with sucrose 5.0 g and then lyophilized. The lyophilized activated oxidized polysaccharide was then reconstituted with DMSO 100 mL at room temperature. The lyophilized CRM 197 (200 mg) was also reconstituted with DMSO 100 mL.

將經氧化之多醣及CRM 197混合，且在室溫下攪拌反應混合物1小時(SPR輸入：1，1 mg/mL反應濃度)。將氰基硼氫化鈉(100 mg/mL於0.15 M SPB中，pH 7.5) 120 µL (1 MEq)添加至反應物中且在室溫下攪拌20小時。在20小時之後，添加氰基硼氫化鈉(100 mg/mL於0.15 M SPB中，pH 7.5) 120 µL (1 MEq)以淬滅，且隨後在室溫下攪拌(200 rpm) 48小時。隨後用800 mL預冷卻的含5 mM丁二酸鹽之生理食鹽水(pH 6.0)稀釋反應混合物，且藉由UF/DF使用300K MWCO RC膜相對於含5 mM丁二酸鹽之生理食鹽水(pH 6.0) (50×透濾體積)進行純化。在UF/DF之後，滲餘物通過0.22 µm過濾器(Millipak 40)進行過濾，且進行分析。 The oxidized polysaccharide and CRM 197 were mixed and the reaction mixture was stirred at room temperature for 1 hour (SPR input: 1, 1 mg/mL reaction concentration). Sodium cyanoborohydride (100 mg/mL in 0.15 M SPB, pH 7.5) 120 µL (1 MEq) was added to the reaction and stirred at room temperature for 20 hours. After 20 hours, sodium cyanoborohydride (100 mg/mL in 0.15 M SPB, pH 7.5) 120 µL (1 MEq) was added to quench and then stirred at room temperature (200 rpm) for 48 hours. The reaction mixture was then diluted with 800 mL of pre-chilled saline (pH 6.0) containing 5 mM succinate and purified by UF/DF using a 300K MWCO RC membrane versus 5 mM succinate saline (pH 6.0) (50× filter volume). After UF/DF, the raffinate was filtered through a 0.22 µm filter (Millipak 40) and analyzed.

4.  在DMSO中使用還原胺化化學方法(RAC) (RAC-DMSO)進行結合，用NaCNBH 3作為還原劑以及用NaBH 4作為封端劑(充分還原，表2之第1行) 將血清型38經氧化之多醣(參見步驟1) (250 mg)與蔗糖6.25 g混合，且隨後凍乾。隨後將凍乾多醣在室溫下用DMSO 125 mL進行復原。凍乾的CRM 197(600 mg)亦用DMSO 240 mL進行復原。 4. Conjugation using reductive amination chemistry (RAC) in DMSO (RAC-DMSO) with NaCNBH 3 as reducing agent and NaBH 4 as capping agent (fully reduced, row 1 of Table 2) Serotype 38 oxidized polysaccharide (see step 1) (250 mg) was mixed with sucrose 6.25 g and then lyophilized. The lyophilized polysaccharide was then reconstituted with DMSO 125 mL at room temperature. Lyophilized CRM 197 (600 mg) was also reconstituted with DMSO 240 mL.

將125 mg經氧化之多醣及157 mg K-CRM混合，且在室溫下攪拌反應混合物1小時(SPR輸入：0.8)。將氰基硼氫化鈉(100 mg/mL於0.15 M SPB中，pH 7.5，老化3小時) 76 µL (1 MEq)添加至反應物中且在室溫下攪拌20小時。在20小時之後，添加硼氫化鈉(100 mg/mL於WFI中) 46 µL (1 MEq)以淬滅反應物且隨後在室溫下攪拌(200 rpm) 3小時。隨後用600 mL預冷卻的含5 mM丁二酸鹽之生理食鹽水(pH 6.0)稀釋反應混合物，且藉由UF/DF使用100K MWCO RC膜相對於含5 mM丁二酸鹽之生理食鹽水(pH 6.0) (50×透濾體積)進行純化。在UF/DF之後，滲餘物通過0.22 µm過濾器(Millipak 40)進行過濾，且進行分析。125 mg of oxidized polysaccharide and 157 mg of K-CRM were mixed and the reaction mixture was stirred at room temperature for 1 hour (SPR input: 0.8). Sodium cyanoborohydride (100 mg/mL in 0.15 M SPB, pH 7.5, aged for 3 hours) 76 µL (1 MEq) was added to the reaction and stirred at room temperature for 20 hours. After 20 hours, sodium borohydride (100 mg/mL in WFI) 46 µL (1 MEq) was added to quench the reaction and then stirred at room temperature (200 rpm) for 3 hours. The reaction mixture was then diluted with 600 mL of pre-chilled saline (pH 6.0) containing 5 mM succinate and purified by UF/DF using a 100K MWCO RC membrane versus saline (pH 6.0) containing 5 mM succinate. After UF/DF, the raffinate was filtered through a 0.22 µm filter (Millipak 40) and analyzed.

下表2包含藉由本發明之方法獲得的血清型38多醣結合物之表徵資料。 表 2 血清型 38 結合物 結合物 # 1 2 3 4 5 天然Poly MW (KDa) 831.5 684 684 684 684 經尺寸設定之Poly MW (KDa) 166 173 173 173 173 化學方法 RAC-DMSO RAC-DMSO RAC-Aq Click Click 用於形成之酮糖(Sug)的還原% 完全還原 約50%還原 約40%還原 未經還原 未經還原 Act poly之活化程度 DO 17 DO 16 DO 16 23% 23% Act Poly Mw (kDa) 157 169 169 220 220 結合物資料 載體蛋白 CRM 197 CRM 197 CRM 197 CRM 197 SCP 結合產率(%) 47 19 55 46 62 SPR (輸出比率) 0.84 0.56 1.42 0.79 0.76 游離醣(%) 8 22 18 20 16 結合物Mw (kDa) 1456 4964 3504 2121 2288 Mw：分子量；SPR：醣與蛋白質比率；NA：不適用；Act Poly：經活化之多醣 Table 2 below contains the characterization data of the serotype 38 polysaccharide conjugate obtained by the method of the present invention. Table 2 Serotype 38 conjugate Conjugate # 1 2 3 4 5 Natural Poly MW (KDa) 831.5 684 684 684 684 Size-set Poly MW (KDa) 166 173 173 173 173 Chemical methods RAC-DMSO RAC-DMSO RAC-Aq Click Click Reduction % of ketose (Sug) formed Fully restored About 50% restored About 40% restored Not restored Not restored Act poly activation degree DO 17 DO 16 DO 16 twenty three% twenty three% Act Poly Mw (kDa) 157 169 169 220 220 Conjugate data Carrier protein CRM 197 CRM 197 CRM 197 CRM 197 SCP Binding yield (%) 47 19 55 46 62 SPR (Speed Flow Ratio) 0.84 0.56 1.42 0.79 0.76 Free sugars (%) 8 twenty two 18 20 16 Conjugate Mw (kDa) 1456 4964 3504 2121 2288 Mw: molecular weight; SPR: carbohydrate to protein ratio; NA: not applicable; Act Poly: activated polysaccharide

實例 7 ： 使用不同化學 方法 及載體蛋白評估經肺炎鏈球菌血清型 38 醣結合物接種之小鼠中的免疫原性 ( 圖 7 )在標準條件下測定了使用不同化學方法(RAC-Aq、RAC-DMSO或點擊)及不同載體蛋白(CRM 197或SCP)產生之小鼠中的血清型38結合物的調理吞噬幾何平均效價(OPA GMT)。 Example 7 : Evaluation of immunogenicity in mice vaccinated with S. pneumoniae serotype 38 glycoconjugates using different chemistries and carrier proteins ( FIG . 7 ) The opsonophagocytic geometric mean titer (OPA GMT) of serotype 38 conjugates in mice produced using different chemistries (RAC-Aq, RAC-DMSO or click) and different carrier proteins (CRM 197 or SCP) was determined under standard conditions.

已測試五種結合物(參見表2中測試結合物之屬性)。Five conjugates were tested (see Table 2 for properties of the conjugates tested).

在第0週，經由皮下途徑用0.01 µg/動物或0.1 µg/動物之測試結合物對每組二十五隻6至8週齡雌性瑞士韋伯斯特小鼠(Swiss Webster mice)進行免疫接種。在第3週用相同劑量之結合物對小鼠進行加強，且隨後在第5週抽血。用100 µg/劑量之AlPO 4作為佐劑來調配各疫苗接種。所有臨床前免疫原性研究均使用每組25隻小鼠以偵測OPA效價的4至5倍差異。在第二次疫苗接種(第5週，PD 2)之後兩週自小鼠收集全血，且使用血清進行分析。對第5週血清樣品進行血清型特異性OPA。 Twenty-five female Swiss Webster mice aged 6 to 8 weeks were immunized subcutaneously at week 0 with 0.01 µg/animal or 0.1 µg/animal of the test conjugate. Mice were boosted with the same dose of the conjugate at week 3 and subsequently bled at week 5. Each vaccination was formulated with 100 µg/dose of AlPO 4 as an adjuvant. All preclinical immunogenicity studies used 25 mice per group to detect 4- to 5-fold differences in OPA titers. Whole blood was collected from mice two weeks after the second vaccination (week 5, PD 2) and serum was used for analysis. Week 5 serum samples were tested for serotype-specific OPA.

使用調理吞噬活性(OPA)檢定來量測鼠類血清中對肺炎鏈球菌血清型38具有特異性之功能性抗體。在檢定反應中設定測試血清，該檢定反應量測莢膜多醣特異性免疫球蛋白調理細菌、觸發補體沉積從而促進吞噬細胞吞噬及殺滅細菌的能力。OPA效價定義為相對於無測試血清之對照孔，引起細菌計數降低50%之倒數稀釋。OPA效價自涵蓋此50%殺滅截止值之兩個稀釋度內插而得到的。The opsonophagocytic activity (OPA) assay is used to measure functional antibodies specific for S. pneumoniae serotype 38 in murine sera. Test sera are set up in an assay reaction that measures the ability of capsular polysaccharide-specific immunoglobulins to opsonize bacteria, trigger complement deposition, and thereby promote phagocytosis and killing of bacteria by phagocytes. The OPA titer is defined as the reciprocal dilution that causes a 50% reduction in bacterial count relative to control wells without test serum. The OPA titer is interpolated from two dilutions that cover this 50% kill cutoff.

OPA程序係基於Hu等人(2005) Clin Diagn Lab Immunol 12 (2):287-295中所描述之方法，且進行以下修改。將測試血清連續稀釋2.5倍且添加至微量滴定檢定盤中。將活血清型38目標細菌菌株添加至孔中且在25℃下振盪盤30分鐘。向孔中添加分化HL-60細胞(吞噬細胞)及幼兔血清(3至4週齡，PEL-FREEZ®，12%最終濃度)，且在37℃下在振盪盤45分鐘。在培育之後，將10 µL等分試樣轉移至含有50 µL水之MULTISCREEN® HTS過濾盤(MILLIPORE®)的孔中。在真空下通過盤過濾液體，且將50 µL之HYSOY®培養基添加至各孔中且過濾。隨後將過濾盤在37℃、5% CO 2下培育隔夜，且隨後用70% EtOH溶液固定。接著，將盤用考馬斯藍染色且脫色一次。在Cellular Technology Limited (CTL) (Shaker Heights, OH) IMMUNOSPOT®分析儀上對菌落進行成像且計數。原始菌落計數用於繪製殺滅曲線且計算OPA效價。 The OPA procedure was based on the method described in Hu et al. (2005) Clin Diagn Lab Immunol 12 (2): 287-295 with the following modifications. Test sera were serially diluted 2.5-fold and added to microtiter assay plates. Live serotype 38 target bacterial strain was added to the wells and the plate was shaken for 30 minutes at 25°C. Differentiated HL-60 cells (phagocytic cells) and baby rabbit serum (3 to 4 weeks old, PEL-FREEZ®, 12% final concentration) were added to the wells and shaken for 45 minutes at 37°C. After incubation, 10 μL aliquots were transferred to wells of MULTISCREEN® HTS filter plates (MILLIPORE®) containing 50 μL of water. The liquid was filtered through the plate under vacuum, and 50 μL of HYSOY® medium was added to each well and filtered. The filter plate was then incubated overnight at 37°C, 5% CO 2 , and then fixed with 70% EtOH solution. The plate was then stained with Coomassie blue and destained once. Colonies were imaged and counted on a Cellular Technology Limited (CTL) (Shaker Heights, OH) IMMUNOSPOT® analyzer. The raw colony counts were used to plot the kill curve and calculate the OPA titer.

表3中顯示了在不同劑量下五週的OPA效價(幾何平均效價(GMT)以及95%信賴區間(CI))。結果亦呈現於圖7中。 表 3 - 使用RAC -Aq 、RAC -DMSO 或點擊化學方法及CRM 197 或SCP 載體蛋白疫苗接種Pn338 結合物之後的OPA 效價。 結合物用於在佐劑存在下對動物進行疫苗接種。雌性瑞士韋伯斯特小鼠，6 至8 週齡 ；劑量：0 .01 及0 .1 μg / 動物 + AlPO 4 ；疫苗接種：第0 週 及第3 週 ；第 5 週最終放血讀數：OPA 0.01 0.1 RAC/DMSO-CRM完全還原(表2之結合物#1) GMT 15 175 無反應者% 68 13 RAC/DMSO-CRM約50%還原(表2之結合物#2) GMT 12 498 無反應者% 75 0 RAC/Aq-CRM約40%還原(表2之結合物#3) GMT 14 4095 無反應者% 67 4 點擊-CRM (表2之結合物#4) GMT 6 77 無反應者% 96 36 點擊-SCP (表2之結合物#5) GMT 53 900 無反應者% 33 0 GMT：幾何平均效價 Table 3 shows the OPA titers (geometric mean titers (GMT) and 95% confidence intervals (CI)) at five weeks at different doses. The results are also presented in Figure 7. Table 3 - OPA titers after vaccination of Pn338 conjugates using RAC -Aq , RAC -DMSO or hit chemistry and CRM 197 or SCP carrier protein vaccines. Conjugates were used to vaccinate animals in the presence of adjuvant. Female Swiss Webster mice, 6 to 8 weeks of age ; Dosage: 0.01 and 0.1 μg / animal + AlPO 4 ; Vaccination: Week 0 and Week 3 ; Final bleeding reading at Week 5 : OPA 0.01 0.1 RAC/DMSO-CRM fully reduced (Conjugate #1 in Table 2) GMT 15 175 Non-responders% 68 13 RAC/DMSO-CRM is reduced by about 50% (Conjugate #2 in Table 2) GMT 12 498 Non-responders% 75 0 RAC/Aq-CRM is reduced by about 40% (conjugate #3 in Table 2) GMT 14 4095 Non-responders% 67 4 Click-CRM (Conjugate #4 in Table 2) GMT 6 77 Non-responders% 96 36 Click-SCP (Conjugate #5 in Table 2) GMT 53 900 Non-responders% 33 0 GMT: Geometric mean titer

實例 8 ： 在水性系統中使用還原胺化化學方法 ( RAC - Aq ) 之 額外肺炎鏈球菌血清型 38 結合物使用RAC-Aq產生額外血清型38結合物(參見實例6)。 Example 8 : Additional S. pneumoniae serotype 38 conjugates using reductive amination chemistry ( RAC - Aq ) in an aqueous system Additional serotype 38 conjugates were generated using RAC-Aq (see Example 6).

下表4包含藉由本發明之方法獲得的血清型38多醣結合物之表徵資料。 表 4 血清型 38 結合物 結合物 # 6 天然Poly MW (KDa) 376 經尺寸設定之Poly MW (KDa) 140 化學方法 RAC-Aq Act poly之活化程度 DO 15 Act Poly Mw (kDa) 149 結合物資料 載體蛋白 CRM 197 結合產率(%) 67 SPR (輸出比率) 1.07 游離醣(%) 29 結合物Mw (kDa) 3722 Mw：分子量；SPR：醣與蛋白質比率；Act Poly：經活化之多醣 Table 4 below contains the characterization data of the serotype 38 polysaccharide conjugate obtained by the method of the present invention. Table 4 Serotype 38 conjugate Conjugate # 6 Natural Poly MW (KDa) 376 Size-set Poly MW (KDa) 140 Chemical methods RAC-Aq Act poly activation degree DO 15 Act Poly Mw (kDa) 149 Conjugate data Carrier protein CRM 197 Binding yield (%) 67 SPR (Speed Flow Ratio) 1.07 Free sugars (%) 29 Conjugate Mw (kDa) 3722 Mw: molecular weight; SPR: carbohydrate to protein ratio; Act Poly: activated polysaccharide

實例 9 ： 使用點擊化學 ( 點擊 ) 之額外肺炎鏈球菌血清型 38 結合物使用點擊產生額外血清型38結合物(參見實例5)。 Example 9 : Additional S. pneumoniae serotype 38 binders using click chemistry ( click ) Additional serotype 38 binders were generated using click (see Example 5).

下表5包含藉由本發明之方法獲得的血清型38多醣結合物之表徵資料。 表 5 血清型 38 結合物 結合物 # 7 8 9 10 經尺寸設定之Poly MW (KDa) 140 211 146 211 經活化之多醣屬性 Act poly之活化程度 12% 12% 20% 20% Act Poly Mw (kDa) 149 194 151 214 經活化之載體屬性 活化程度(DoA) 20 20 20 20 經活化之載體MW, kDa 104.1 104.1 104.1 104.1 結合物資料 載體蛋白 SCP SCP SCP SCP 結合產率(%) 56 91 73 92 SPR (輸出比率) 0.63 0.99 0.91 0.97 游離醣(%) 15 17 16 11 結合物Mw (kDa) 1061 5611 3382 9598 Mw：分子量；SPR：醣與蛋白質比率；NA：不適用；Act Poly：經活化之多醣 Table 5 below contains the characterization data of the serotype 38 polysaccharide conjugate obtained by the method of the present invention. Table 5 Serotype 38 conjugate Conjugate # 7 8 9 10 Size-set Poly MW (KDa) 140 211 146 211 Properties of activated polysaccharides Act poly activation degree 12% 12% 20% 20% Act Poly Mw (kDa) 149 194 151 214 Properties of activated carriers Degree of Activation (DoA) 20 20 20 20 Activated carrier MW, kDa 104.1 104.1 104.1 104.1 Conjugate data Carrier protein SCP SCP SCP SCP Binding yield (%) 56 91 73 92 SPR (Speed Flow Ratio) 0.63 0.99 0.91 0.97 Free sugars (%) 15 17 16 11 Conjugate Mw (kDa) 1061 5611 3382 9598 Mw: molecular weight; SPR: carbohydrate to protein ratio; NA: not applicable; Act Poly: activated polysaccharide

實例 10 ： 使用不同化學方法及載體蛋白評估經肺炎鏈球菌血清型 38 醣結合物接種之小鼠中的免疫原性 ( 圖 8 )在標準條件下測定了使用不同化學方法(RAC-Aq或點擊)及不同載體蛋白(CRM 197或SCP)產生之小鼠中的血清型38結合物的調理吞噬幾何平均效價(OPA GMT)。 Example 10 : Evaluation of immunogenicity in mice vaccinated with S. pneumoniae serotype 38 glycoconjugates using different chemistries and carrier proteins ( FIG. 8 ) The opsonophagocytic geometric mean titer (OPA GMT) of serotype 38 conjugates in mice produced using different chemistries (RAC-Aq or hit) and different carrier proteins (CRM 197 or SCP) was determined under standard conditions.

已測試七種結合物(參見表2、表4及表5中測試結合物之屬性)。Seven conjugates were tested (see Tables 2, 4 and 5 for properties of the tested conjugates).

在第0週，經由皮下途徑用0.01 µg/動物或0.1 µg/動物之測試結合物對每組二十五隻6至8週齡雌性瑞士韋伯斯特小鼠進行免疫接種。在第3週用相同劑量之結合物對小鼠進行加強，且隨後在第5週抽血。用100 µg/劑量之AlPO 4作為佐劑來調配各疫苗接種。所有臨床前免疫原性研究均使用每組25隻小鼠以偵測OPA效價的4至5倍差異。在第二次疫苗接種(第5週，PD 2)之後兩週自小鼠收集全血，且使用血清進行分析。對第5週血清樣品進行血清型特異性OPA。 Twenty-five female Swiss Webster mice aged 6 to 8 weeks were immunized subcutaneously at week 0 with 0.01 µg/animal or 0.1 µg/animal of the test conjugate. Mice were boosted with the same dose of the conjugate at week 3 and subsequently bled at week 5. Each vaccination was formulated with 100 µg/dose of AlPO 4 as an adjuvant. All preclinical immunogenicity studies used 25 mice per group to detect 4- to 5-fold differences in OPA titers. Whole blood was collected from mice two weeks after the second vaccination (week 5, PD 2) and serum was used for analysis. Week 5 serum samples were tested for serotype-specific OPA.

使用調理吞噬活性(OPA)檢定來量測鼠類血清中對肺炎鏈球菌血清型38具有特異性之功能性抗體(參見實例7)。The opsonophagocytic activity (OPA) assay was used to measure functional antibodies specific for S. pneumoniae serotype 38 in mouse sera (see Example 7).

表6中顯示了在不同劑量下五週的OPA效價(幾何平均效價(GMT)以及95%信賴區間(CI))。結果亦呈現於圖8中。 表 6 - 使用RAC -Aq 或點擊化學方法及CRM 197 或SCP 載體蛋白疫苗接種Pn338 結合物之後的OPA 效價。 結合物用於在佐劑存在下對動物進行疫苗接種。雌性瑞士韋伯斯特小鼠 ，6 至8 週齡； 劑量 ：0 .01 及0 .1 μg / 動物 + AlPO 4 ； 疫苗接種 ： 第0 週 及第3 週； 第5 週最終放血讀數 ：OPA 0.01 0.1 RAC/Aq-CRM (表2之結合物#3) GMT NA 3334 無反應者% NA 4 RAC/Aq-CRM (表4之結合物#6) GMT NA 1023 無反應者% NA 4 點擊-SCP (表2之結合物#5) GMT 40 663 無反應者% 48 8 點擊-SCP (表5之結合物#7) GMT 9 74 無反應者% 79 28 點擊-SCP (表5之結合物#9) GMT 113 264 無反應者% 24 16 點擊-SCP (表5之結合物#10) GMT 29 869 無反應者% 44 0 點擊-SCP (表5之結合物#8) GMT 18 1002 無反應者% 56 4 NA：不適用 GMT：幾何平均效價 Table 6 shows the OPA titers (geometric mean titers (GMT) and 95% confidence intervals (CI)) at five weeks at different doses. The results are also presented in Figure 8. Table 6 - OPA titers after vaccination with Pn338 conjugates using RAC -Aq or hit chemistry and CRM 197 or SCP carrier protein vaccines . Conjugates were used to vaccinate animals in the presence of adjuvant. Female Swiss Webster mice , 6 to 8 weeks of age; Dosage : 0.01 and 0.1 μg / animal + AlPO 4 ; Vaccination : Week 0 and Week 3; Final bleeding reading at Week 5 : OPA 0.01 0.1 RAC/Aq-CRM (Conjugate #3 in Table 2) GMT NA 3334 Non-responders% NA 4 RAC/Aq-CRM (Conjugate #6 in Table 4) GMT NA 1023 Non-responders% NA 4 Click-SCP (Conjugate #5 of Table 2) GMT 40 663 Non-responders% 48 8 Click-SCP (Conjugate #7 of Table 5) GMT 9 74 Non-responders% 79 28 Click-SCP (Conjugate #9 of Table 5) GMT 113 264 Non-responders% twenty four 16 Click-SCP (Conjugate #10 of Table 5) GMT 29 869 Non-responders% 44 0 Click-SCP (Conjugate #8 of Table 5) GMT 18 1002 Non-responders% 56 4 NA: Not Applicable GMT: Geometric Mean Titer

實例 11 ： 使用點擊化學 ( 點擊 ) 之額外肺炎鏈球菌血清型 38 結合物已使用點擊產生額外血清型38結合物(參見實例5)，然而使用碳酸N,N'-二丁二醯亞胺酯(DSC) (1.5 MEq)代替CDI。表7示出了表徵資料。 表 7 血清型 38 結合物 結合物 # 11 經尺寸設定之Poly MW (KDa) 202 經活化之多醣屬性 Act poly之活化程度 30% Act Poly Mw (kDa) 246 結合物資料 載體蛋白 SCP 結合產率(%) 83 SPR (輸出比率) 0.8 游離醣(%) ＞5 結合物Mw (kDa) 5291 Mw：分子量；SPR：醣與蛋白質比率；NA：不適用；Act Poly：經活化之多醣 Example 11 : Additional S. pneumoniae serotype 38 conjugates using click chemistry ( click ) Additional serotype 38 conjugates have been generated using click (see Example 5), however N,N'-disuccinimidyl carbonate (DSC) (1.5 MEq) was used instead of CDI. Table 7 shows the characterization data. Table 7 Serotype 38 conjugate Conjugate # 11 Size-set Poly MW (KDa) 202 Properties of activated polysaccharides Act poly activation degree 30% Act Poly Mw (kDa) 246 Conjugate data Carrier protein SCP Binding yield (%) 83 SPR (Speed Flow Ratio) 0.8 Free sugars (%) >5 Conjugate Mw (kDa) 5291 Mw: molecular weight; SPR: carbohydrate to protein ratio; NA: not applicable; Act Poly: activated polysaccharide

本說明書中提及之所有公開案及專利申請案指示熟習本發明所屬領域之技術者的水平。所有公開案及專利申請案均在此以引用之方式併入，如同各個別公開案或專利申請案特定且單獨地指示為以引用之方式併入一般。All publications and patent applications mentioned in this specification are indicative of the level of skill in the art to which the invention pertains. All publications and patent applications are hereby incorporated by reference as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.

儘管出於清晰理解的目的已經藉助於說明及實例相當詳細地描述了前述發明，但可在隨附申請專利範圍之範疇內實踐某些改變及修改。Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, certain changes and modifications may be practiced within the scope of the appended claims.

圖 1A 及圖 1B.肺炎鏈球菌多醣血清型38重複單元組織之示意圖，包括最常見形式(絲胺酸形式) (A)及二級形式(甘胺酸形式) (B)。 圖 2.肺炎鏈球菌血清型38-絲胺酸形式之結構在水合物與酮狀態之間形成平衡。兩種狀態由黑色圓圈顯示。 圖 3.O-乙醯化(下圖)及去O-乙醯化(上圖)血清型38多醣(絲胺酸形式)之擴展變旋異構及甲基區域1D 1H光譜。針對兩個光譜標註變旋異構及甲基信號。陰影框表示由於β-D-Gal f殘基之O-乙醯化而觀測到的共振，且不存在於去O-乙醯化血清型38多醣中。 圖 4.O-乙醯化(下圖)及去O-乙醯化(上圖)血清型38多醣(絲胺酸形式)之擴展羰基、變旋異構及甲基區域1D 13C光譜。圖A至圖D分別展示了酮、羰基、變旋異構及甲基共振之擴展區。陰影框表示由於β-D-Galp殘基之O-乙醯化而觀測到的共振，且不存在於去O-乙醯化血清型38多醣中。 圖 5.經還原之血清型38多醣的1D 1H譜。(插圖顯示擴展的變旋異構區)。標註變旋異構及甲基信號。 圖 6.用NaBH 4還原之前及之後的血清型38重複單元(絲胺酸形式)之示意圖。還原形成FucpNAc及QuipNAc糖。 圖 7.來自使用RAC/水溶液、RAC/DMSO或點擊結合，用與CRM 197或SCP結合之血清型38多醣進行接種之小鼠血清的Pn38特異性調理吞噬幾何平均效價(OPA GMT)。 圖 8.來自使用RAC/水溶液或點擊結合，用與CRM 197或SCP結合之血清型38多醣進行接種之小鼠血清的Pn38特異性調理吞噬幾何平均效價(OPA GMT)。 Figure 1A and Figure 1B. Schematic representation of the organization of the repeat unit of the S. pneumoniae polysaccharide serotype 38, including the most common form (serine form) (A) and the secondary form (glycine form) (B). Figure 2. The structure of the S. pneumoniae serotype 38-serine form in equilibrium between the hydrate and keto states. The two states are shown by black circles. Figure 3. Expanded mutameric and methyl region 1D 1 H spectra of O-acetylated (lower panel) and de-O-acetylated (upper panel) serotype 38 polysaccharide (serine form). Mutameric and methyl signals are annotated for both spectra. The shaded boxes represent the resonances observed due to O-acetylation of the β-D-Gal f residue and are not present in the de-O-acetylated serotype 38 polysaccharide. Figure 4. 1D 13 C spectra of the expanded carbonyl, isomeric and methyl regions of O-acetylated (lower panel) and de-O-acetylated (upper panel) serotype 38 polysaccharide (serine form). Panels A to D show the expanded regions of keto, carbonyl, isomeric and methyl resonances, respectively. The shaded boxes represent the resonances observed due to O-acetylation of the β-D-Gal p residue and are not present in the de-O-acetylated serotype 38 polysaccharide. Figure 5. 1D 1 H spectrum of reduced serotype 38 polysaccharide. (Inset shows expanded mutameric region). Mutameric and methyl signals are annotated. Figure 6. Schematic representation of serotype 38 repeat units (serine form) before and after reduction with NaBH4 . Reduction forms FucpNAc and QuipNAc sugars. Figure 7. Pn38-specific opsonophagocytic geometry mean titers (OPA GMT) from sera from mice vaccinated with serotype 38 polysaccharides conjugated to CRM 197 or SCP using RAC/aqueous solution, RAC/DMSO, or click-binding. Figure 8. Pn38-specific opsonophagocytic geometry mean titers (OPA GMT) from sera from mice vaccinated with serotype 38 polysaccharides conjugated to CRM 197 or SCP using RAC/aqueous solution or click-binding.

TW202444421A_113111933_SEQL.xmlTW202444421A_113111933_SEQL.xml

Claims (34)

一種醣結合物，其包含具有以下重複單元之經分離之肺炎鏈球菌(S. pneumoniae)血清型38醣： 其中n表示重複單元之數目，其中Y表示絲胺酸或甘胺酸殘基，其中所有該等重複單元包含相同的Y殘基，且其中在β-D-Gal p4OAc,6(Y)之位置4處的O-乙醯基存在於約0%至約100%的該等重複單元中，該醣與載體蛋白結合。 A carbohydrate conjugate comprising an isolated Streptococcus pneumoniae (S. pneumoniae) serotype 38 saccharide having the following repeating units: wherein n represents the number of repeat units, wherein Y represents a serine or glycine residue, wherein all of the repeat units comprise the same Y residue, and wherein the O-acetyl group at position 4 of β-D-Gal p 4OAc,6(Y) is present in about 0% to about 100% of the repeat units, and the sugar is bound to a carrier protein. 如請求項1之血清型38醣結合物，其中該血清型38醣結合物係藉由點擊化學製備。The serotype 38 glycoconjugate of claim 1, wherein the serotype 38 glycoconjugate is prepared by click chemistry. 一種血清型38醣結合物，其包含具有以下重複單元之血清型38莢膜醣： 其中n表示重複單元之數目，其中Y表示絲胺酸或甘胺酸殘基，其中所有該等重複單元包含相同的Y殘基，且其中X表示N-乙醯基-D-岩藻糖胺(N-acetyl-D-fucosamine，D-FucNAc)殘基或N-乙醯基-D-奎諾糠胺(N-acetyl-D-quinovosamine，D-QuiNAc)殘基。 A serotype 38 glycoconjugate comprising a serotype 38 capsular saccharide having the following repeating units: wherein n represents the number of repeating units, wherein Y represents a serine or glycine residue, wherein all of the repeating units contain the same Y residue, and wherein X represents an N-acetyl-D-fucosamine (D-FucNAc) residue or an N-acetyl-D-quinovosamine (D-QuiNAc) residue. 一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(IV)： ， 其中X係選自由CH 2(CH 2) n '、(CH 2CH 2O) mCH 2CH 2、NHCO(CH 2) n '、NHCO(CH 2CH 2O) mCH 2CH 2、OCH 2(CH 2) n '及O(CH 2CH 2O) mCH 2CH 2組成之群；其中n'係選自1至10且m係選自1至4， 其中X'係選自由CH 2O(CH 2) n''CH 2C=O、CH 2O(CH 2CH 2O) m'(CH 2) n''CH 2C=O組成之群，其中n''係選自0至10且m'係選自0至4， 其中方括號中之結構表示該血清型38醣之重複單元，且其中n表示重複單元之數目。 A serotype 38 saccharide conjugate comprises a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and has the general formula (IV): , wherein X is selected from the group consisting of CH 2 (CH 2 ) n ′ , (CH 2 CH 2 O) m CH 2 CH 2 , NHCO(CH 2 ) n ′ , NHCO(CH 2 CH 2 O) m CH 2 CH 2 , OCH 2 (CH 2 ) n ′ , and O(CH 2 CH 2 O) m CH 2 CH 2 ; wherein n′ is selected from 1 to 10 and m is selected from 1 to 4, wherein X′ is selected from the group consisting of CH 2 O(CH 2 ) n′ CH 2 C═O, CH 2 O(CH 2 CH 2 O) m′ (CH 2 ) n′ CH 2 C═O, wherein n′ is selected from 0 to 10 and m′ is selected from 0 to 4, wherein the structure in square brackets represents the repeating unit of the serotype 38 carbohydrate, and wherein n represents the number of repeating units. 一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(IV)，其中X為CH 2(CH 2) n '，其中n'為2，且其中X'為CH 2O(CH 2) n ''CH 2C=O，其中n''為1。 A serotype 38 saccharide conjugate comprises a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and has the general formula (IV), wherein X is CH 2 (CH 2 ) n ′ , wherein n′ is 2, and wherein X′ is CH 2 O(CH 2 ) n ′′ CH 2 C═O, wherein n′′ is 1. 一種血清型38醣結合物，其包含透過間隔子與載體蛋白(CP)共價結合之血清型38醣且具有通式(V)， 其中方括號中之結構表示該血清型38醣之重複單元，且其中n表示重複單元之數目。 A serotype 38 saccharide conjugate comprises a serotype 38 saccharide covalently bound to a carrier protein (CP) via a spacer and has the general formula (V), Wherein the structure in square brackets represents the repeating unit of the serotype 38 carbohydrate, and wherein n represents the number of repeating units. 如請求項1至6中任一項之醣結合物，其中在結合之前該肺炎鏈球菌血清型38醣之重量平均分子量(Mw)在150 kDa與300 kDa之間。The saccharide conjugate of any one of claims 1 to 6, wherein the weight average molecular weight (Mw) of the Streptococcus pneumoniae serotype 38 saccharide before conjugation is between 150 kDa and 300 kDa. 如請求項1至7中任一項之醣結合物，其中該血清型38醣結合物之重量平均分子量(Mw)在1,000 kDa與10,000 kDa之間。The glycoconjugate of any one of claims 1 to 7, wherein the weight average molecular weight (Mw) of the serotype 38 glycoconjugate is between 1,000 kDa and 10,000 kDa. 如請求項1至8中任一項之醣結合物，其中在結合之前該肺炎鏈球菌血清型38醣之重量平均分子量(Mw)在150 kDa與300 kDa之間，且其中該血清型38醣結合物之重量平均分子量(Mw)在1,000 kDa與10,000 kDa之間。The glycoconjugate of any one of claims 1 to 8, wherein the weight average molecular weight (Mw) of the Streptococcus pneumoniae serotype 38 saccharide prior to conjugation is between 150 kDa and 300 kDa, and wherein the weight average molecular weight (Mw) of the serotype 38 saccharide conjugate is between 1,000 kDa and 10,000 kDa. 如請求項1至9中任一項之醣結合物，其中該血清型38醣結合物之結合程度在2與15之間。The glycoconjugate of any one of claims 1 to 9, wherein the degree of binding of the serotype 38 glycoconjugate is between 2 and 15. 如請求項1至10中任一項之醣結合物，其中該血清型38醣結合物之結合程度在4與10之間。The glycoconjugate of any one of claims 1 to 10, wherein the degree of binding of the serotype 38 glycoconjugate is between 4 and 10. 如請求項1至6中任一項之醣結合物，其中在結合之前該肺炎鏈球菌血清型38醣之重量平均分子量(Mw)在150 kDa與300 kDa之間，其中該血清型38醣結合物之重量平均分子量(Mw)在1,000 kDa與10,000 kDa之間且其中該血清型38醣結合物之結合程度在4與10之間。The glycoconjugate of any one of claims 1 to 6, wherein the weight average molecular weight (Mw) of the Streptococcus pneumoniae serotype 38 saccharide prior to conjugation is between 150 kDa and 300 kDa, wherein the weight average molecular weight (Mw) of the serotype 38 glycoconjugate is between 1,000 kDa and 10,000 kDa and wherein the degree of conjugation of the serotype 38 glycoconjugate is between 4 and 10. 如請求項1至12中任一項之醣結合物，其中該醣結合物中之血清型38醣與載體蛋白之比率(w/w)在0.5與1.5之間。The glycoconjugate of any one of claims 1 to 12, wherein the ratio (w/w) of serotype 38 saccharide to carrier protein in the glycoconjugate is between 0.5 and 1.5. 如請求項1至12中任一項之醣結合物，其中該醣結合物中之血清型38醣與載體蛋白之比率(w/w)在0.7與1.1之間。The glycoconjugate of any one of claims 1 to 12, wherein the ratio (w/w) of serotype 38 saccharide to carrier protein in the glycoconjugate is between 0.7 and 1.1. 如請求項1至6中任一項之醣結合物，其中在結合之前該肺炎鏈球菌血清型38醣之重量平均分子量(Mw)在150 kDa與300 kDa之間，其中該血清型38醣結合物之重量平均分子量(Mw)在1,000 kDa與10,000 kDa之間，其中該醣結合物中之血清型38醣與載體蛋白之比率(w/w)在0.5與1.5之間且其中該血清型38醣結合物之結合程度在4與10之間。The glycoconjugate of any one of claims 1 to 6, wherein the weight average molecular weight (Mw) of the Streptococcus pneumoniae serotype 38 saccharide before conjugation is between 150 kDa and 300 kDa, wherein the weight average molecular weight (Mw) of the serotype 38 glycoconjugate is between 1,000 kDa and 10,000 kDa, wherein the ratio (w/w) of serotype 38 saccharide to carrier protein in the glycoconjugate is between 0.5 and 1.5 and wherein the degree of conjugation of the serotype 38 glycoconjugate is between 4 and 10. 如請求項1至6中任一項之醣結合物，其中在結合之前該肺炎鏈球菌血清型38醣之重量平均分子量(Mw)在150 kDa與300 kDa之間，其中該血清型38醣結合物之重量平均分子量(Mw)在1,000 kDa與10,000 kDa之間，其中該醣結合物中之血清型38醣與載體蛋白之比率(w/w)在0.7與1.1之間且其中該血清型38醣結合物之結合程度在4與10之間。The glycoconjugate of any one of claims 1 to 6, wherein the weight average molecular weight (Mw) of the Streptococcus pneumoniae serotype 38 saccharide before conjugation is between 150 kDa and 300 kDa, wherein the weight average molecular weight (Mw) of the serotype 38 glycoconjugate is between 1,000 kDa and 10,000 kDa, wherein the ratio (w/w) of serotype 38 saccharide to carrier protein in the glycoconjugate is between 0.7 and 1.1 and wherein the degree of conjugation of the serotype 38 glycoconjugate is between 4 and 10. 如請求項1至16中任一項之醣結合物，其中該血清型38醣結合物包含與血清型38醣之總量相比小於約20%之游離血清型38醣。The carbohydrate conjugate of any one of claims 1 to 16, wherein the serotype 38 carbohydrate conjugate comprises less than about 20% free serotype 38 carbohydrate compared to the total amount of serotype 38 carbohydrate. 如請求項1至6中任一項之醣結合物，其中在結合之前該肺炎鏈球菌血清型38醣之重量平均分子量(Mw)在150 kDa與300 kDa之間，其中該血清型38醣結合物之重量平均分子量(Mw)在1,000 kDa與10,000 kDa之間，其中該醣結合物中之血清型38醣與載體蛋白之比率(w/w)在0.5與1.5之間，其中該血清型38醣結合物之結合程度在4與10之間且其中該血清型38醣結合物包含與血清型38醣之總量相比小於約20%之游離血清型38醣。The glycoconjugate of any one of claims 1 to 6, wherein the weight average molecular weight (Mw) of the Streptococcus pneumoniae serotype 38 saccharide prior to conjugation is between 150 kDa and 300 kDa, wherein the weight average molecular weight (Mw) of the serotype 38 saccharide conjugate is between 1,000 kDa and 10,000 kDa, wherein the ratio (w/w) of serotype 38 saccharide to carrier protein in the glycoconjugate is between 0.5 and 1.5, wherein the degree of conjugation of the serotype 38 saccharide conjugate is between 4 and 10, and wherein the serotype 38 saccharide conjugate comprises less than about 20% of free serotype 38 saccharide compared to the total amount of serotype 38 saccharide. 如請求項1至6中任一項之醣結合物，其中在結合之前該肺炎鏈球菌血清型38醣之重量平均分子量(Mw)在150 kDa與300 kDa之間，其中該血清型38醣結合物之重量平均分子量(Mw)在1,000 kDa與10,000 kDa之間，其中該醣結合物中之血清型38醣與載體蛋白之比率(w/w)在0.7與1.1之間，其中該血清型38醣結合物之結合程度在4與10之間且其中該血清型38醣結合物包含與血清型38醣之總量相比小於約20%之游離血清型38醣。The glycoconjugate of any one of claims 1 to 6, wherein the weight average molecular weight (Mw) of the Streptococcus pneumoniae serotype 38 saccharide prior to conjugation is between 150 kDa and 300 kDa, wherein the weight average molecular weight (Mw) of the serotype 38 saccharide conjugate is between 1,000 kDa and 10,000 kDa, wherein the ratio (w/w) of serotype 38 saccharide to carrier protein in the glycoconjugate is between 0.7 and 1.1, wherein the degree of conjugation of the serotype 38 saccharide conjugate is between 4 and 10, and wherein the serotype 38 saccharide conjugate comprises less than about 20% of free serotype 38 saccharide compared to the total amount of serotype 38 saccharide. 如請求項1至19中任一項之醣結合物，其中至少60%的該血清型38醣結合物在CL-4B管柱中之K d小於或等於0.3。 The glycoconjugate of any one of claims 1 to 19, wherein at least 60% of the serotype 38 glycoconjugate has a K d of less than or equal to 0.3 in a CL-4B column. 如請求項1至19中任一項之醣結合物，其中65%與80%之間的該血清型38醣結合物在CL-4B管柱中之K d低於或等於0.3。 The glycoconjugate of any one of claims 1 to 19, wherein between 65% and 80% of the serotype 38 glycoconjugate has a K d of less than or equal to 0.3 in a CL-4B column. 如請求項1至6中任一項之醣結合物，其中在結合之前該肺炎鏈球菌血清型38醣之重量平均分子量(Mw)在150 kDa與300 kDa之間，其中該血清型38醣結合物之重量平均分子量(Mw)在1,000 kDa與10,000 kDa之間，其中該醣結合物中之血清型38醣與載體蛋白之比率(w/w)在0.5與1.5之間，其中該血清型38醣結合物之結合程度在4與10之間，其中該血清型38醣結合物包含與血清型38醣之總量相比小於約20%之游離血清型38醣且其中65%與80%之間的該血清型38醣結合物在CL-4B管柱中之K d低於或等於0.3。 The glycoconjugate of any one of claims 1 to 6, wherein the weight average molecular weight (Mw) of the Streptococcus pneumoniae serotype 38 saccharide before conjugation is between 150 kDa and 300 kDa, wherein the weight average molecular weight (Mw) of the serotype 38 saccharide conjugate is between 1,000 kDa and 10,000 kDa, wherein the ratio (w/w) of serotype 38 saccharide to carrier protein in the glycoconjugate is between 0.5 and 1.5, wherein the degree of conjugation of the serotype 38 saccharide conjugate is between 4 and 10, wherein the serotype 38 saccharide conjugate comprises less than about 20% of free serotype 38 saccharide compared to the total amount of serotype 38 saccharide and wherein between 65% and 80% of the serotype 38 saccharide conjugate has a Kd in a CL-4B column of less than or equal to 0.3. 如請求項1至6中任一項之醣結合物，其中在結合之前該肺炎鏈球菌血清型38醣之重量平均分子量(Mw)在150 kDa與300 kDa之間，其中該血清型38醣結合物之重量平均分子量(Mw)在1,000 kDa與10,000 kDa之間，其中該醣結合物中之血清型38醣與載體蛋白之比率(w/w)在0.7與1.1之間，其中該血清型38醣結合物之結合程度在4與10之間，其中該血清型38醣結合物包含與血清型38醣之總量相比小於約20%之游離血清型38醣且其中65%與80%之間的該血清型38醣結合物在CL-4B管柱中之K d低於或等於0.3。 The glycoconjugate of any one of claims 1 to 6, wherein the weight average molecular weight (Mw) of the Streptococcus pneumoniae serotype 38 saccharide prior to conjugation is between 150 kDa and 300 kDa, wherein the weight average molecular weight (Mw) of the serotype 38 saccharide conjugate is between 1,000 kDa and 10,000 kDa, wherein the ratio (w/w) of serotype 38 saccharide to carrier protein in the glycoconjugate is between 0.7 and 1.1, wherein the degree of conjugation of the serotype 38 saccharide conjugate is between 4 and 10, wherein the serotype 38 saccharide conjugate comprises less than about 20% of free serotype 38 saccharide compared to the total amount of serotype 38 saccharide and wherein between 65% and 80% of the serotype 38 saccharide conjugate has a Kd in a CL-4B column of less than or equal to 0.3. 如請求項1至23中任一項之醣結合物，其中該血清型38醣結合物之該載體蛋白係選自由以下組成之群：TT、DT、DT突變體及來自鏈球菌(Streptococcus)之C5a肽酶(SCP)。The glycoconjugate of any one of claims 1 to 23, wherein the carrier protein of the serotype 38 glycoconjugate is selected from the group consisting of TT, DT, DT mutants and C5a peptidase (SCP) from Streptococcus. 如請求項1至23中任一項之醣結合物，其中該血清型38醣結合物之該載體蛋白係根瘤菌抗生物素蛋白(rhizavidin) [aa 45-179J-GGGGSSS-SP1500- AAA-SP0785] (CP1)。The glycoconjugate of any one of claims 1 to 23, wherein the carrier protein of the serotype 38 glycoconjugate is rhizavidin [aa 45-179J-GGGGSSS-SP1500-AAA-SP0785] (CP1). 如請求項1至23中任一項之醣結合物，其中該血清型38醣結合物之該載體蛋白為Rhavi-連接子-PdT(G294P)-連接子-SP0435 [aa 62-185]融合蛋白(SPP2)。The glycoconjugate of any one of claims 1 to 23, wherein the carrier protein of the serotype 38 glycoconjugate is Rhavi-linker-PdT(G294P)-linker-SP0435 [aa 62-185] fusion protein (SPP2). 如請求項1至23中任一項之醣結合物，其中該血清型38醣結合物之該載體蛋白為SCP。The glycoconjugate of any one of claims 1 to 23, wherein the carrier protein of the serotype 38 glycoconjugate is SCP. 一種免疫原組合物，其包含如請求項1至27中任一項之肺炎鏈球菌血清型38醣結合物。An immunogenic composition comprising the Streptococcus pneumoniae serotype 38 glycoconjugate of any one of claims 1 to 27. 如請求項28之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型1、2、3、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15B、18C、19A、19F、22F、23F及33F之醣結合物。The immunogenic composition of claim 28, further comprising a carbohydrate conjugate from Streptococcus pneumoniae serotypes 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F and 33F. 如請求項28之免疫原組合物，其進一步包含來自肺炎鏈球菌血清型1、3、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15A、15B、18C、19A、19F、22F、23A、23B、23F、24F、33F及35B之醣結合物。The immunogenic composition of claim 28, further comprising a saccharide conjugate from Streptococcus pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F and 35B. 如請求項30之免疫原組合物，其中來自血清型1、4、5、6A、6B、7F、8、9V、10A、11A、12F、14、15A、15B、18C、19A、19F、22F、23A、23B、23F、24F、33F、35B及38之該肺炎鏈球菌醣與CRM 197結合且該肺炎鏈球菌醣血清型3與SCP結合。 The immunogenic composition of claim 30, wherein the pneumococcal saccharides from serotypes 1, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, 35B and 38 are bound to CRM 197 and the pneumococcal saccharide serotype 3 is bound to SCP. 如請求項28至31中任一項之免疫原組合物，其進一步包含一種佐劑。The immunogenic composition of any one of claims 28 to 31, further comprising an adjuvant. 如請求項28至32中任一項之免疫原組合物，其用作疫苗。An immunogenic composition according to any one of claims 28 to 32, for use as a vaccine. 如請求項28至32中任一項之免疫原組合物，其用於預防、治療或改善個體之與肺炎鏈球菌血清型38相關的感染、疾病或病況的方法中。An immunogenic composition as claimed in any one of claims 28 to 32, for use in a method of preventing, treating or ameliorating an infection, disease or condition associated with Streptococcus pneumoniae serotype 38 in a subject.
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